Automated data storage system

Abstract

A block for data storage. [Solution] A system for data storage is described, comprising components for receiving, encoding, synthesizing, storing, reading, sequencing, and decoding digital information stored in polynucleotides and libraries of polynucleotides.

Description

[Technical Field]

[0001] Cross-reference of related applications

[0001] This application claims the benefits and priority of U.S. Provisional Patent Application No. 63 / 492,593 filed on 28 March 2023 and U.S. Provisional Patent Application No. 63 / 514,227 filed on 18 July 2023, the contents of which are incorporated herein by reference in their entirety. All publications, patents, and patent applications referenced herein are incorporated by reference to the same extent as each individual publication, patent, or patent application is specifically and individually indicated as being incorporated by reference. [Background technology]

[0002] background

[0002] DNA is an attractive data storage medium given its superior density, stability, energy efficiency, and lifespan compared to current storage methods. However, automated systems may involve processes that perform several different components, such as coding, writing / synthesis, storage, and reading / sequencing. Therefore, there is a need to develop systems that can perform these functions. [Overview of the project] [Means for solving the problem]

[0003] overview

[0003] This specification provides a system for data storage comprising a computing system comprising at least one processor and instructions executable by the at least one processor to perform one or more operations, and a modular rack-mount synthesizer. In some embodiments, one or more operations include receiving digital information; encoding digital information in one or more nucleic acid sequences; and synthesizing a library of polynucleotides corresponding to nucleic acid sequences. In some embodiments, the modular rack-mount synthesizer unit includes a computer controller, one or more reservoirs, at least one flow cell block including one or more flow cells, an extraction stage, a post-processing unit, a storage unit, a robotic interfering unit, and / or a rack interface. In some embodiments, the at least one flow cell block includes at least 12 flow cells. In some embodiments, one or more flow cells include one or more solid supports configured to synthesize a library of polynucleotides; and one or more ports for exchanging gases, synthesis reagents, and / or extracted polynucleotides.

[0004]

[0004] In some embodiments, the modular rack-mount synthesis unit includes an extraction stage and / or a post-processing unit, the extraction stage and / or post-processing unit including an energy supply chain, one or more ports for exchanging synthesis reagents and / or extracted polynucleotides. In some embodiments, the modular rack-mount synthesis unit includes a storage unit, the storage unit comprising a storage plate, a robotic interface configured to move the storage plate, and an access port. In some embodiments, the modular rack-mount synthesis unit includes a rack interface, the rack interface comprising a power supply, a rack reagent partition, a synthesis unit reagent partition, one or more reservoirs, an energy supply chain, and a pilot valve bank. In some embodiments, the storage unit provides a library of polynucleotides to a sequencing unit configured to sequence the library of polynucleotides.

[0005]

[0005] In some embodiments, the system for data storage further comprises one or more sensors configured to detect a state, the state including temperature, pressure, humidity, voltage changes, current changes, capacitance, conductivity, storage plate position, liquid volume, flow rate, and / or presence of liquid, and to report the state to a computing system. In some embodiments, a modular rack-mount synthesis unit comprises at least one flow cell block, and one or more sensors further configured to pause synthesis in one or more flow cells or at least one flow cell block. In some embodiments, the system is a first system for data storage, and the computing system is configured to route coded data from (i) a paused flow cell or (ii) a paused flow cell block to an active flow cell, an active flow cell block, or a second system for data storage.

[0006]

[0006] In some embodiments, the computing system further includes a cache where digital information is held in the cache until polynucleotides encoding the digital information are stored.

[0007]

[0007] Furthermore, the Specified Specification provides an assembly for storing information, comprising: a plurality of compartments comprising a first material, each compartment configured to receive a plurality of polynucleotides encoding information; a base plate comprising a second material; and a cover plate comprising a third material, wherein the coefficient of thermal expansion (CTE) of the first material, the second material, or both is smaller than that of the third material. In some embodiments, the plurality of compartments are arranged in an array on the base plate. In some embodiments, the base plate, the cover plate, or both include a plurality of recessed features, and each compartment is located at least partially within a recessed feature. In some embodiments, the first material comprises a borosilicate. In some embodiments, the second material has a specific heat capacity of about 0.5 J / k-°C to 2.5 J / k-°C. In some embodiments, the third material includes a CTE of about 15 μm / m-℃ to 20 μm / m-℃; a thermal conductivity of about 15 W / mK to about 20 W / mK; and / or a specific heat capacity of about 0.5 J / g-℃. In some embodiments, the third material includes stainless steel.

[0008] Brief explanation of the drawing

[0008] A better understanding of the features and advantages of the methods and apparatus described herein will be obtained by referring to the following detailed description and accompanying drawings that describe exemplary embodiments. [Brief explanation of the drawing]

[0009] [Figure 1A]

[0009] This is a flowchart illustrating an exemplary workflow of an automated data storage system using nucleic acids including a synthetic module according to an aspect of the present disclosure. [Figure 1B]

[0010] This flowchart shows the workflow of an automated data storage system using nucleic acids, including a storage module. In some examples, these steps are performed by robotic stages according to aspects of this disclosure. [Figure 1C]

[0011] This flowchart shows an exemplary system architecture workflow at the server rack level for an automated data storage system (as described herein) using nucleic acids, including firmware, a control system (such as a network interface controller), and a virtual data path, according to aspects of this disclosure. In some aspects, data may move to a server rack for storage and verification, and the server rack may transmit data to an external interface for database management. [Figure 2A]

[0012] This figure shows an exemplary standardized DNA synthesis unit (DSU) according to aspects of the present disclosure, including data / addressing, power, and reagent / DNA input / removal (flow cell, flow cell block, or entire data storage unit), and includes a table indicating the dimensions and chip density of an exemplary 6U module. [Figure 2B]

[0013] This is a system-level diagram illustrating an exemplary automated data storage system using nucleic acids. It shows modules for reagents, rack mounting, synthesis, post-processing (e.g., deprotection, drying, PCR, purification), control, power distribution, and status indicators according to aspects of this disclosure. [Figure 3A]

[0014] This figure shows an exemplary rack unit for data storage, displaying a DNA data storage system unit extending from a rack unit according to an aspect of the present disclosure. [Figure 3B]

[0015] This figure shows the mechanical layout of the DNA data storage system unit shown in Figure 3A, according to an aspect of this disclosure. [Figure 3C]

[0016] This figure shows a schematic front / transparent view of an exemplary DNA data storage system unit configured for use in a rack unit as shown in Figure 3A, according to an aspect of the present disclosure. [Figure 3D]

[0017] This figure shows a schematic diagram of the back panel of an exemplary DNA data storage system unit according to an aspect of the present disclosure. [Figure 4A]

[0018] Two figures illustrate the mechanical layout of an extraction stage of an exemplary DNA data storage system unit according to aspects of this disclosure. The top figure shows an extraction stage in which the storage chamber is located in a first position for distribution at the front of the stage. The bottom figure shows an extraction stage in which the storage chamber is moved to a second position at the rear of the stage for waste / washing. These two positions are shown for illustrative purposes only, and in some examples the chamber may be moved to any position associated with the flow cell block. [Figure 4B]

[0019] This figure shows a side view of the fluid pathway of an exemplary DNA data storage system unit, including reagent delivery, according to an aspect of the present disclosure. [Figure 4C]

[0020] This figure shows a top view of the fluid pathway of an exemplary DNA data storage system unit, including reagent delivery, according to an aspect of the present disclosure. [Figure 4D]

[0021] This figure shows a schematic diagram of the fluid path in an exemplary flow cell block, including a control valve. [Figure 4E]

[0022] This figure shows an exemplary flow cell block device illustrating fluid input. Twelve flow cell units are shown as an example only. [Figure 5A]

[0023] Three diagrams illustrating an exemplary flow cell block for synthesis, including a filling, washing, and drying process using a Barkart valve (6712 series, 24V), according to aspects of the present disclosure. [Figure 5B]

[0024] Three diagrams illustrating an exemplary flow cell block for extraction, including a washing, filling, and extraction process using a Barcart valve (6712 series, 24V), according to aspects of the present disclosure. [Figure 5C]

[0025] This figure shows a schematic diagram of an exemplary flow cell block for extraction according to an aspect of the present disclosure, in which angles alpha (α) and beta (β) are labeled, and the x, y, and z axes are labeled. [Figure 6]

[0026] This is a piping and instrumentation diagram illustrating an exemplary 12-flow cell unit design according to aspects of the present disclosure. Each flow cell is indicated by an inlet and an outlet. The inlet supplies reagents for biomolecule synthesis. The outlet provides a fluid path for waste and may be placed under positive pressure from a gas source (e.g., nitrogen). The inlet may also be bypassed for biomolecule extraction. [Figure 7]

[0027] This figure shows a schematic of the process for liquid-phase deprotection of nucleic acids in the device described herein, according to aspects of the present disclosure. The process comprises one or more of the following steps: loading a chip into a flow cell (FC); filling the flow cell with a deprotection solution; closing a valve and incubating for a period of time; opening the valve and collecting the liquid containing the polynucleotide; incubating in AMA (ammonium hydroxide / 40% aqueous methylamine 1:1 v / v) at 65°C for 20 minutes, or incubating with TBA (tert-butylamine); and drying the tube. [Figure 8]

[0028] This figure shows a schematic representation of a nucleic acid data storage system according to an aspect of this disclosure, having modules for input, synthesis, storage, amplification / processing, and sequencing, which are operated by a controller and a robotic system. In some aspects, the systems described herein include one or more of these types of units. [Figure 9A]

[0029] Figure 9A shows an exemplary structure for storing multiple polynucleotides according to an aspect of this disclosure. The structure used for nucleic acid storage may be stored on a plate or other device described herein. Figure 9A shows a substantially tubular structure. [Figure 9B]

[0029] Figure 9B shows an exemplary structure for storing multiple polynucleotides according to an aspect of the present disclosure. Structures used for nucleic acid storage may be stored on a plate or other devices described herein. Figure 9B shows a structure including a cap and a body that are flush-welded together. [Figure 9C]

[0029] Figure 9C shows an exemplary structure for storing multiple polynucleotides according to an aspect of the present disclosure. Structures used for nucleic acid storage may be stored on a plate or other devices described herein. Figure 9C shows a structure including a removable screw cap. [Figure 9D]

[0029] Figure 9D shows an exemplary structure for storing multiple polynucleotides according to an aspect of the present disclosure. Structures used for nucleic acid storage may be stored on a plate or other device described herein. Figure 9D shows a structure including a septum. [Figure 9E]

[0029] Figure 9E shows an exemplary structure for storing multiple polynucleotides according to an aspect of the present disclosure. Structures used for nucleic acid storage may be stored on a plate or other devices described herein. Figure 9E shows a structure comprising two round, tablet-shaped hemispheres that form a seal when one half is inserted into the other half. [Figure 9F]

[0029] Figure 9F shows an exemplary structure for storing multiple polynucleotides according to an aspect of the present disclosure. Structures used for nucleic acid storage may be stored on a plate or other devices described herein. Figure 9F shows a structure comprising a substantially flat disk-shaped chamber with a sealable lid. [Figure 9G]

[0029] Figure 9G shows an exemplary structure for storing multiple polynucleotides according to an aspect of the present disclosure. Structures used for nucleic acid storage may be stored on a plate or other devices described herein. Figure 9G shows a structure including a box with an optionally fitted lid. [Figure 9H]

[0029] Figure 9H shows an exemplary structure for storing multiple polynucleotides according to an aspect of the present disclosure. Structures used for nucleic acid storage may be stored on a plate or other devices described herein. Figure 9H shows a structure including a surface radio frequency identification (RFID) tag. [Figure 9I]

[0029] Figure 9I shows an exemplary structure for storing multiple polynucleotides according to an aspect of the present disclosure. Structures used for nucleic acid storage may be stored on a plate or other devices described herein. Figure 9I shows a structure including an embedded RFID tag. [Figure 10]

[0030] This figure shows an exemplary computing device having one or more processors, memory, storage, and network interfaces according to an aspect of the present disclosure. [Figure 11]

[0031] This is a flowchart illustrating an exemplary method for storing digital information in multiple polynucleotides according to an aspect of the present disclosure. [Figure 12]

[0032] This flowchart shows an exemplary method for obtaining digital information in multiple polynucleotides according to an aspect of the present disclosure. [Figure 13]

[0033] This figure shows an exemplary passive RFID system according to the embodiments of this disclosure. [Figure 14]

[0034] This figure shows a non-limiting example of digital information divided into multiple sub-items for storage in a structure, according to the aspects of this disclosure. [Figure 15A]

[0035] Figure 15A is a side view of an exemplary compartment for storing polynucleotides according to an aspect of the present disclosure. Figure 15A shows a first set of dimensions of the compartment using a pipette to indicate filling or retrieving polynucleotides in solution. [Figure 15B]

[0035] Figure 15B is a side view of an exemplary compartment for storing polynucleotides according to an aspect of the present disclosure. Figure 15B shows the same side view as Figure 15A, but with further dimensions not shown in Figure 15A. [Figure 16]

[0036] This is a diagram illustrating an exemplary cover plate or base plate for a system for storing polynucleotides according to an aspect of the present disclosure. [Figure 17]

[0037] This diagram shows exemplary cover plates before (solid lines) and after (dotted lines) exposure to heat, according to aspects of the present disclosure. In some aspects, the expansion of the cover plates is exaggerated for illustrative purposes. [Figure 18]

[0038] This is a schematic diagram showing a cross-section of an exemplary assembly for storing polynucleotides according to an aspect of the present disclosure. The assembly includes a base plate, compartments for holding polynucleotides, and a cover plate. In some embodiments, the wall angles of the cover plate wall and / or base plate wall are exaggerated for illustrative purposes, and the wall angles of the cover plate wall and / or base plate wall may be less than 0.5°. [Figure 19]

[0039] This is a table reporting the properties of various types of glass (GA-1, GA-4, GA-9, GA-12, GA-13, GA-21, GA-34, GA-44, and GA-47) that may be selected for compartments for storing polynucleotides, in accordance with aspects of this disclosure. [Figure 20]

[0040] This flowchart shows an exemplary method for sealing multiple polynucleotides in a storage device according to an aspect of the present disclosure. [Modes for carrying out the invention]

[0010] Detailed description

[0041] Data Storage Gym

[0042] Currently available data storage technologies based on hard disk drives and tape drives face significant physical, technical, and investment challenges. This has led to a substantial slowdown in the rate of increase in storage capacity, and coupled with the need to migrate data every 5-10 years (depending on the storage medium), it has a significant impact on the cost of long-term (over 10 years) storage. Furthermore, the ratio of long-term ("cold") data to active ("warm" / "hot") data is dramatically increasing, and combined with the slowdown in the increase of data density, this is putting pressure on the utilization, footprint, and storage capacity of data center infrastructure. In contrast, the systems, devices, and methods provided herein that utilize DNA molecules as a storage medium result in improvements in both data longevity / retention time and volumetric storage density.

[0011]

[0043] In addition, to ensure the stability and viability of DNA used for long-term data storage (e.g., over 30 years), it may be important that the encapsulation method protects the encapsulated DNA from external contamination. This includes not only the ingress of water, but also other harmful gases and liquids. Current methods can generally be aimed at long-term DNA storage on a scale of several millennia, for example, by encapsulating dried DNA in borosilicate glass inserts sealed within small metal shells and laser welding them in an inert atmosphere. Accelerated aging experiments have shown that the half-life of DNA stored under ambient conditions can reach as high as 38,000 years.

[0012]

[0044] However, while currently available methods and systems may be used for long-term DNA data storage in archives due to the expected lifespan of the stored DNA, they have many drawbacks for use in long-term enterprise or hyperscale archive storage applications, including, for example, data center / rack-scale applications. Specifically, not only are the materials themselves expensive, but the sealing techniques may involve a high-power (YAG) laser for laser welding the top of the capsule and associated robotic automation systems for positioning and arranging individual capsules to facilitate the welding process. These processes can be cumbersome, expensive, unreliable, require high maintenance, have slow throughput, and / or occupy a very large amount of space. Furthermore, the capsules storing the DNA itself can be relatively large, and importantly, considerably larger than the mass of DNA they contain, thereby significantly reducing the achievable volumetric storage capacity. Due to these constraints, these systems cannot compete with the volumetric storage capacity of conventional data tapes (e.g., LTO 10 cartridges). The systems and methods described herein can overcome such limitations while retaining the advantages of long-term storage.

[0013]

[0045] Provided herein are data storage systems that utilize a highly modularized synthesis, processing, and storage framework. In some examples, such systems are based on standard sizes. In some examples, such systems are based on the OCP (Open Computing) data center rack form factor, which simplifies data center utilization and adoption. In some examples, such systems are based on the standard OCP (21-inch / 48OU) data center rack form factor, which simplifies data center utilization and adoption. In some examples, such systems are based on the standard (19-inch / 48U) data center rack form factor, which simplifies data center utilization and adoption. Further provided herein are systems incorporating a number of tens to hundreds of individual synthesis / flow cells integrated into a single, remotely serviced synthesis module. In some examples, a central robotic system is used to position a storage plate containing tens to hundreds of automatically filled storage chambers (or containers, e.g., wells or capsules) to access the DNA synthesized by each individual flow cell. In some examples, the robotic system then moves the filled plates to additional modules in storage racks for subsequent post-processing steps, including but not limited to intermediate dry-down, PCR, dispensing for quality control and monitoring, final dry-down for storage, and plate sealing. In some examples, the robotic system moves the processed and sealed plates to storage modules for long-term storage (e.g., up to several decades). In some examples, the systems provided herein interface with magnetic or solid-state memory systems. In some examples, the data storage system includes one or more components shown in Figure 8.This specification provides a computing system for data storage, comprising at least one processor and instructions executable by at least one processor for performing operations, wherein the operations include receiving digital information, encoding digital information into one or more nucleic acid sequences, and synthesizing a library of polynucleotides corresponding to nucleic acid sequences, and a system comprising one or more modular rack-mount synthesis units. In some examples, the data storage system includes one or more of a computer controller, a flow cell block, an extraction stage, a post-processing unit, a storage unit, a robotic interface, and a rack interface.

[0014]

[0046] Furthermore, this specification provides systems and methods for encapsulation. In some examples, the systems and methods described herein utilize hermetic sealing methods, and in some examples, are specifically applied to DNA data storage. In some examples, the systems and methods described herein reduce the complexity of the sealing mechanism compared to those currently available. In some examples, the encapsulation volume is also reduced by using small compartments, such as vials encapsulated within a thin metal cover plate, for example, in some embodiments. Furthermore, in some examples, high mechanical rigidity and / or a very thin profile / form factor for the final assembly is provided by adding a metal base plate made of the same or similar material as the cover plate. In some examples, the method includes hermetic sealing under an inert atmosphere. In some examples, the hermetic sealing method includes a glass-metal hermetic sealing method. Glass-metal sealing can generally be used to create a robust feedthrough to a vacuum-sealed package. Glass-metal sealing can generally be achieved by two different mechanisms, namely compression sealing or fit sealing. Compression sealing can utilize a mismatch in the coefficient of thermal expansion (CTE) between the glass insert and the outer housing. In some examples, a seal can be established. This is because, as the components cool after heat treatment, thermal compression of the outer housing acts on the glass insert. Housing materials with high CTE, such as stainless steel or titanium, may be suitable for compression seals. Depending on the design, compression seals can withstand very high levels of pressure (e.g., up to 3000 bar), and this type of glass-metal seal may be suitable for microelectronics connectors, including ultra-high vacuum windows, components used in spacecraft, high-pressure sensors for industrial and oil and gas applications, and defense applications, among other uses.

[0015]

[0047] This specification provides devices or assemblies for storing information. In some examples, an assembly for storing information may have one or more of a plurality of compartments, a base plate, and at least one cover plate. In some examples, the plurality of compartments contain a first material. In some examples, the base plate contains a second material. In some examples, at least one cover plate contains a third material. In some examples, a method for storing information is provided herein. In some examples, a method for storing information includes one or more of providing an assembly having a plurality of compartments, a base plate, and a cover plate, and generating a temperature gradient between the base plate and the cover plate. In some examples, the plurality of compartments contain a plurality of polynucleotides. In some examples, the temperature gradient causes the base plate, the cover plate, or both to expand or contract to seal the plurality of compartments. In some examples, a system for storing information is provided herein. In some examples, a system for storing information includes one or more of an assembly for storing information, a material deposition system, one or more temperature control systems, and a computing system. In some examples, the assembly includes one or more of several compartments, a base plate, and at least one cover plate. In some examples, the material deposition system includes a dispenser. In some examples, the dispenser deposits multiple polynucleotides into one of the several compartments. In some examples, one or more temperature control systems are used to heat or cool the base plate, the cover plate, or both. In some examples, the computing system includes at least one processor and instructions executable by at least one processor to perform one or more operations. In some examples, one or more operations include coordinating the movement of one or more components of the system, monitoring the properties of one or more components of the system, or both.

[0016]

[0048] Methods and systems for tracking content within a DNA data storage system are further provided herein. Digital information may be encoded into multiple polynucleotides stored in a structure of the DNA data storage system. A structure may include tags for identifying the structure, tags for providing metadata about the content within the structure, or a combination thereof. Tags can be used as labels, file systems, or a combination thereof. For example, tags can be used to remotely catalog structures within a data storage system, enabling individual identification and selection of structures from the data storage system. Tags can also provide data fixation because they can provide physically directly associated information about the content of the structure.

[0017]

[0049] In some examples, the system stores digital information. In some examples, the system includes multiple polynucleotides that collectively encode the digital information. In some examples, the system further includes a structure for storing multiple polynucleotides. In some examples, the structure includes a radio frequency identification (RFID) tag. In some examples, the RFID tag includes metadata about the multiple polynucleotides. In some examples, the method provides for storing digital information. In some examples, the method includes synthesizing multiple polynucleotides. In some examples, the multiple polynucleotides collectively encode the digital information. In some examples, the method includes writing metadata about the multiple polynucleotides to a radio frequency identification (RFID) tag. In some examples, the method includes storing multiple polynucleotides in a structure. In some examples, the structure includes an RFID tag. In some examples, the method determines the data integrity of the stored digital information. In some examples, the method includes writing metadata about the multiple polynucleotides to a radio frequency identification (RFID) tag. In some examples, the method includes scanning the RFID tag after a duration to determine data integrity. In some examples, the RFID tag is valid or invalid. In some examples, what is provided herein is a product. In some examples, the product includes a structure for storing multiple polynucleotides that encode digital information. In some examples, the structure includes a radio frequency identification (RFID) tag. In some examples, the RFID tag includes metadata about the multiple polynucleotides.

[0018]

[0050] Data storage unit

[0051] This specification provides data storage systems. In some examples, the data storage system comprises the system shown in Figure 2A. In some examples, the system includes one or more ports for reagent / gas / waste exchange, interfaces for data processing / communication, and multiple solid supports for power and biomolecular synthesis. In some examples, the data storage system comprises one or more modules of the system shown in Figure 2B. The exemplary systems provided herein include, in some examples, one or more of a reagent reservoir, a well plate reservoir, a waste container (or reservoir), and a rack unit, as well as one or more modules. In some examples, the modules include, but are not limited to, synthesis, deprotection, storage, post-processing (after synthesis or after cleavage from solid supports, e.g., deprotection, dry-down, amplification, purification, or other processes), a control module, a power distribution module, and a status indicator (e.g., a panel). In some examples, the rack interface comprises one or more (local) reservoirs. In some examples, the reservoirs are configured to hold waste (organic or aqueous), phosphoramidites, amplification reagents (e.g., PCR or other amplification techniques), and beads.

[0019]

[0052] The data storage system may include 300 (Figure 3A) units. In some examples, the biomolecules include nucleic acids (e.g., DNA, RNA), peptides, sugars, or other biopolymers. In some examples, the data storage system comprises data storage units 303. In some examples, the data storage units are configured to be mounted in rack units 301 (Figures 3A-3C). In some examples, the rack unit includes multiple “slots” 302 into which the data storage units are inserted. In some examples, the data storage units occupy one or more slots within the rack unit. In some examples, the rack unit 301 is used as part of a large-scale data storage center that includes hundreds or even thousands of rack units to store large amounts of data. In some examples, multiple data storage units enable data redundancy. In some examples, the rack unit includes multiple data storage units, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more than 10 data storage units. In some examples, the data storage units provided herein are highly modular, allowing for easy replacement of the entire data storage unit or its components, such as flow cell blocks, flow cells, storage media, or solid supports for biomolecular synthesis. In some examples, the rack-mount data storage units provided herein are compatible with standard (19-inch / 48U) rack units. In some examples, the rack-mount data storage units provided herein are compatible with standard OCP (21-inch / 48OU) rack units.

[0020]

[0053] Rack Interface

[0054] The data storage unit may be configured to interface with the rack unit. In some examples, the data storage unit is configured to be movable within the rack unit (examples of an "open" data storage unit are shown in Figures 3A–3C). In some examples, chains, guides, slides, tracks, rollers, cables, magnets, or other mechanical means are used to allow the data storage system to slide laterally away from the rack unit (e.g., "open") and laterally back into the rack unit (e.g., "closed"). In some examples, this allows user access (e.g., manual user access or robotic system access) to one or more components of the data storage system. In some examples, this allows the data storage unit to be completely removed from the rack. In some examples, removing the data storage unit from the rack unit does not require the use of additional tools or the removal of rivets, screws, or bolts. In some examples, a user or robotic system removes a faulty storage device from the rack and replaces it with a functional storage device. In some examples, the rack unit includes fans, filters, or other means for extracting smoke generated within the system.

[0021]

[0055] In some examples, the rack interface provides power, communication, gas supply, reagent and waste removal, access to specific components of the data storage unit, and performs other functions required for data storage. In some examples, the interface comprises one or more connectors (or ports). In some examples, the connectors or ports are configured to modularly remove the entire synthesis unit or various components of the synthesis unit from the rack. In some examples, the rack interface supplies power, provides communication, supplies gas and reagents, and removes waste from a main reservoir outside the rack unit (e.g., reagent bulkhead 305 / 307). In some examples, the interface includes one or more e-chains (e.g., 306 / 309) configured to deliver power, reagents, gas, or remove waste to various components of the data storage unit. In some examples, the rack interface comprises one or more panels. In some examples, the panels are located in any part of the system, including the top, bottom, side, front, or rear. In some examples, the rack interface includes a front panel and a rear panel. In some examples, the rear panel interface includes one or more connectors and ports as shown in Figure 3D (305 / 307). In some examples, one or more panels are connected via wiring or piping (e.g., two partitions such as 305 and 307). In some examples, the panel includes an access window 318. In some examples, the access window is configured to remove one or more storage plates or chambers from the storage unit. In some examples, the panel includes one or more connectors for performing diagnostics on the storage unit. In some examples, the front panel includes one or more gauges, dials, indicator lights, LCD displays, or other indicators of the status or performance of the data storage unit. In some examples, the panel indicates the status of one or more sensors.One or more sensors may include, in non-limiting examples, a liquid volume sensor, capacitance sensor, temperature sensor, humidity sensor, pressure sensor, salinity sensor, light sensor, UV sensor, conductivity sensor, or gas sensor (e.g., an O2 or N2 sensor). In some examples, the sensors are incorporated into one or more flow cells. In some examples, the sensors are configured to identify reagents or liquids within the flow cell. In some examples, the sensors include an out-of-plane cathode within the flow cell. In some examples, the panel indicates the presence of a fault condition, such as power loss, low gas pressure, and leaking fluid. In some examples, the panel includes a control panel for a module or unit of the system described herein. In some examples, the panel includes an interface that includes a visual indicator of the condition and / or fault indication. In some examples, the fault condition is associated with a fault detected by one or more sensors. In some examples, the panel includes connectors for one or more of the following: extraction, amidite, bulk reagent, waste, power, communication, and gas. In some examples, the gas includes nitrogen or argon. In some examples, at least two, three, four, or at least five different gas pressures are delivered through one or more connectors on the panel. In some examples, the rack interface includes one or more partitions. In some examples, the communication interface includes a wireless communication interface.

[0022]

[0056] In some examples, the interface includes a power connector. In some examples, the power connector supplies approximately 110 or 220 volts to the rack unit. In some examples, the rack unit further includes a transformer 304 (or power supply) for supplying 24V to the composite unit. In some examples, the rack interface includes one or more wire harnesses. In some examples, the wire harnesses are configured as an e-chain. In some examples, the wire harnesses provide communication and / or power to various components of the data storage unit, such as circuit boards, valves, sensors, robots, pumps, or other components. In some examples, the interface further includes a battery backup system. In some examples, the backup system is activated when power is lost to the entire rack or to one or more data storage units within the rack. In some examples, power loss is reported to a central controller, processor, display panel, or remote workstation. In some examples, the communication includes information for the operation of the data storage unit. In some examples, the communication includes digital information encoded in biomolecules.

[0023]

[0057] The rack interface may include one or more e-chains for distributing electricity, liquids, and / or gases. In some examples, e-chains are configured for the delivery of reagents and / or the removal of waste 306. In some examples, e-chains are configured for the extraction of biomolecules from the synthesis unit or storage stage 309. In some examples, e-chains are connected to ports or connectors on the rear panel. In some examples, e-chains are connected to one or more reservoirs. In some examples, the reservoirs include a reagent reservoir. In some examples, the reservoirs include an extraction biomolecule reservoir. In some examples, the reservoirs include a waste reservoir. In some examples, the e-chains include an extraction stage e-chain 309. In some examples, e-chains are shielded or protected from electrical short circuits, interference, or contact with solvents used during biomolecule synthesis. In some examples, e-chains allow the data storage unit to be extended laterally or away from the rack unit without interrupting electricity, reagents, waste, or gases. In some examples, fluids and / or gases are delivered throughout the data storage unit using piping. In some examples, the tubing includes tubes with an ID of approximately 1 / 64 inch, 1 / 32 inch, 1 / 16 inch, or 1 / 8 inch.

[0024]

[0058] The rack interface may include a valve bank 308. In some examples, the valve bank includes a pilot valve bank. In some examples, the valve bank includes at least 4, 8, 12, 16, 24, 36, 48, or at least 64 channels. In some examples, the valve bank includes 4, 8, 12, 16, 24, 36, 48 or fewer, or 64 or fewer channels. In some examples, the valve bank includes 4-64, 4-48, 4-32, 8-64, 8-48, 8-48, 8-32, 8-24, 12-36, 12-48, 12-64, 16-36, 16-64, 16-48, 20-30, 24-36, 24-48, 24-64, or 48-64 channels.

[0025]

[0059] The rack interface may include an Open Compute (OCP) rack design. In some examples, the system includes OCP specification V1.2 or V2.0. In some examples, the Open Compute design features a 21-inch standard with a width of 600 mm. In some examples, the OCP has a 12V / 48V DC power supply. In some examples, the OCP includes one or more DC busbars. In some examples, the busbars are rear-mounted DC busbars. In some examples, the OCP has up to three 12V DC busbars, each of which can be further horizontally divided into two power zones. In some examples, outputs of 6.6kW and 13.2kW are provided over an overall range of up to 40kW. In some examples, the OCP has a 48V DC power supply, making up to 36kW of electrical output available in each rack. In some examples, the OCP server hardware is powered via end-to-end DC busbars in each rack. In some examples, the rack includes an OCP power pack. In some examples, the power pack is configured to generate the required DC from a three-phase AC source. In some examples, power packs are mounted in racks to power data storage systems. These power packs, in some examples, support modular rack mounting and monitoring via a network (SNMP). In some examples, individual power pack blocks generate a typical 3kW output and are connected in parallel. In some examples, the OCP features a battery backup unit integrated into the OCP rack to provide backup power.

[0026]

[0060] System Operations

[0061] This specification provides data storage systems configured to perform one or more methods (or operations) described herein. In some examples, the data storage system includes one or more modules or components configured to perform the methods described herein. In some examples, the system comprises one or more modules provided in Figure 8.

[0027]

[0062] A computer interface may be used to control the inputs to a data storage system provided herein. In some examples, the computer interface is configured to receive data (e.g., users, customers, servers, computers, other data storage systems, or other sources). In some examples, the computer interface is controlled by a user. In some examples, the computer interface is fully automated without substantial human intervention. In some examples, the computer interface includes formatting or transforming input data. In some examples, the computer interface includes encoding, encrypting, decrypting, or deciphering data. In some examples, the computer interface provides output containing instructions (e.g., formatted data) for the synthesis of biomolecules (e.g., nucleic acids). In some examples, the computer interface provides synthesis instructions for one or more synthesis units.

[0028]

[0063] The data storage systems provided herein, in some examples, include an interface for receiving reagent information. In some examples, the reagent information includes the age of the reagent, storage conditions, quantity (e.g., volume), or other data. In some examples, the interface is configured to receive information from one or more sensors located in or near a reservoir. In some examples, one or more reservoirs are configured to store reagents or waste. In some examples, the interface receives information about the reagent quality of regenerated / recovered reagents. In some examples, the interface provides output to a server, cloud, user, or display panel on the system.

[0029]

[0064] The synthesis unit can convert formatted data into one or more biomolecules. In some examples, the formatted data includes in silico biopolymer sequences. In some examples, the biopolymer sequences include nucleic acid sequences. In some examples, the synthesis unit is configured to receive commands for synthesis via a communication port. In some examples, the commands include programs for controlling valves, temperature, humidity, pumps, monitoring sensors, programs for applying power to one or more components of the synthesis unit, or other commands related to the synthesis of biomolecules. In some examples, the synthesis unit provides quality control information about the synthesis process as output. In some examples, the synthesis unit provides information about the synthesized molecules as output storage information, such as memory structure, flow cell identification information, flow cell block identification, total operating time, or other information about the biomolecules synthesized by the synthesis unit. In some examples, the synthesis unit receives commands to cleave or remove biomolecules from one or more solid supports. In some examples, the synthesis unit receives commands to transfer the cleaved biomolecules to an extraction unit. In some examples, all biomolecules are cleaved from one or more solid supports. In some examples, a certain percentage of biomolecules are cleaved from one or more solid supports. In some examples, at least 5%, 10%, 15%, 20%, 25%, 50%, 75%, 90%, 95%, or at least 99% of the biomolecules were cleaved. Exemplary operations for synthesis are shown in Figure 5A. In some examples, the operations control the fluid communication between the solvent, reagents, and gases and the synthesis surface on the solid support 402. In the filling step, the flow cell 401 in the flow cell block 311 is filled with reagents by opening the first valve 501. Any liquid or gas present in the flow cell from the previous step is moved to a liquid waste (e.g., a reservoir) by opening the second valve 502 (Figure 5A, left). The third valve 503 remains closed. In the washing step, similar valves are opened and closed, but instead, a washing solvent flows through the system (Figure 5A, center).In the drying process, a fourth valve 504 connected to the gas is used to push any fluid from the flow cell 401 into the liquid waste via the first valve 501 (Figure 5A, right). The third valve 503 remains closed. In the washing process, liquid is pumped into the waste from the first valve 501 through the third valve 503 (Figure 5B, left). The second valve 502 remains closed. During the extraction process, a fourth valve 504 connected to the gas is used to push any fluid from the flow cell 401 into the extraction chamber via the third valve 503 (Figure 5A, right). The first valve 501 remains closed.

[0030]

[0065] The extraction unit 316 may receive biomolecules synthesized from the synthesis unit. In some examples, the extraction unit is configured to perform one or more post-processing steps after synthesis. Post-processing steps may include, but are not limited to, concentration, drying, amplification, purification, quality control / analysis, cleavage, ligation, selection / capture, deprotection, or other post-processing steps. In some examples, the extraction unit receives instructions to perform a drying step on the biomolecules after synthesis. In some examples, the extraction unit receives instructions to perform an amplification step on the biomolecules after synthesis. In some examples, the extraction unit receives instructions to store the biomolecules in a storage unit. In some examples, the extraction unit receives instructions to test the quality or quantity of the synthesized molecules. In some examples, the extraction unit is configured to perform one or more steps shown in Figure 7. In some examples, the extraction unit is configured to fill a flow cell containing a solid support for synthesis with a deprotection solution; close one or more valves and incubate the flow cell; open one or more valves and recover the extracted biomolecules; and optionally deprotect the biomolecules after extraction.

[0031]

[0066] A storage module (or unit) may be configured to receive extracted biomolecules. In some examples, the storage unit receives instructions for the storage location of biomolecules from a specific synthesis unit 303, flow cell block 311, or flow cell 401. In some examples, the storage unit includes a tray or surface for biomolecular storage 317. In some examples, the storage unit receives instructions to operate a drying unit. As output, in some examples, the storage unit provides the stored biomolecules in the form of chambers (or containers), such as trays, plates, tapes, capsules, or any combination thereof. In some examples, the storage accepts instructions to seal the biomolecules within the trays, plates, tapes, or capsules. In some examples, the storage unit provides metadata specific to one or more chambers as output. In some examples, the storage unit provides metadata specific to one or more capsules as output. In some examples, the storage unit provides output metadata specific to one or more trays. In some examples, the storage unit is configured to perform the steps shown in Figure 1B. In some examples, the storage unit comprises one or more robotic stages 316 for performing one or more of the following steps: tray loading / initialization, moving trays to a synthesis / extraction unit, positioning and moving intermediate trays, centrifugal separation of trays under low temperature or vacuum conditions, moving trays to a sealing station, and moving trays to a storage module (e.g., Figure 4A). In some examples, the storage unit comprises a stage 316 configured to move one or more storage chambers to different flow cell locations in a synthesis unit. In some examples, the storage unit includes multiple redundant data copies stored on the same plate or in the same location (e.g., a well) across multiple locations. In some examples, one or more storage units are configured in a RAID-type storage configuration. In some examples, instance RAID 0, RAID 1, RAID 5, RAID 6, or RAID 10 configurations are used.In some examples, the configuration includes one or more of striping, mirroring, and parity (dispersion or dual parity), including any combination thereof. In some examples, the storage unit includes plates. In some examples, the plates include multiple wells. In some examples, the number of wells is proportional to the number of flow cell blocks or flow cells. In some examples, the storage device includes wells corresponding to each flow cell in a flow cell block. In some examples, the storage module includes a deprotection module. In some examples, the deprotection module is configured to perform the operations shown in Figure 2B, including desealing, incubation, and resealing. In some examples, the storage module includes a drydown module. In some examples, the drydown module includes a vacuum / centrifuge, evaporator, or other system for drying biomolecules.

[0032]

[0067] A quality control unit may be used to evaluate synthesized biomolecules. In some examples, the quality control unit includes one or more spectroscopic devices, sequencers, or other systems for evaluating biomolecules. In some examples, the quality control unit accepts one or more extracted biomolecules as input. In some examples, the quality control unit provides a log file containing biomolecular information as output. In some examples, the biomolecular information includes sequence, concentration, quantity, or other information.

[0033]

[0068] controller

[0069] The devices, systems, or platforms provided herein for biomolecular extraction may be integrated into a data storage system. The data storage system may include one or more modules. In some examples, some or all of the modules are interconnected. In some examples, some or all of the modules are interconnected, allowing for the transfer of polynucleotides between them. In some examples, some or all of the modules are fluidically coupled. In some examples, some or all of the modules are fluidically coupled to one or more tubes. The fluid generally refers, without limitation, to one or more liquids used in various processes involved in handling polynucleotides, including synthesis, amplification, sequencing preparation, and sequencing. In some examples, some or all of the modules are interconnected, allowing for the transfer of control commands between modules in the system. In some examples, some or all of the modules are electronically coupled. Modules in the system may include, without limitation, synthesizer units, amplification chambers, sequencer units, storage units, controllers, robotic systems, or any combination thereof. In some examples, modules may further include a fluid source, a database or file system, or both. In some examples, a database or file system records the system's storage capacity. For example, the database or file system may record the available racks (or trays), slots (for capsules), or both. In some examples, the database or file system is used to determine the placement of racks within the storage system. In some examples, the movement of polynucleotides between one or more modules of the system is achieved by one or more tube or robotic systems. In some examples, the database or file system is used to direct robotic systems to the correct locations within the storage system. In some examples, the system is autonomous. In some examples, a controller system is configured to perform operations between modules, as shown in Figure 1C.

[0034]

[0070] A system for storing information may include a device or assembly for storing information, such as a device including a plurality of compartments, a base, a plate, and / or a cover plate, as further described herein. The system may further include a material deposition system including a dispenser, as described herein. The dispenser may be used to deposit a plurality of polynucleotides into one of the plurality of compartments. The dispenser may be used to deposit reagents for processes described herein, such as the synthesis, amplification, and / or sequencing of polynucleotides. The system may further include one or more temperature control systems. One or more temperature control systems may be used to heat or cool one or more components of the device or system, such as a base plate, a cover plate, or a plurality of compartments or a portion thereof. The system may further include a computing system, as described herein. The computing system may comprise at least one processor and instructions executable by at least one processor to perform one or more operations. In some examples, one or more operations include coordinating the movement of one or more components of the system and / or monitoring the properties of one or more components of the system. In some examples, coordinating the movement of one or more components of a system includes positioning a cover plate on a base plate, placing a cover plate on a base plate, or both. In some examples, coordinating the movement of one or more components of a system includes aligning a dispenser in a material deposition system into one of several compartments. In some examples, the properties include temperature, humidity, pressure, salinity, a photosensor, UV, O2, or any combination thereof. In some examples, monitoring the properties of one or more components includes monitoring the temperature of components in a device within a system, such as a polynucleotide storage device including a base plate, a cover plate, or multiple components including polynucleotides.

[0035]

[0071] A data storage system may include a control system. Generally, a control system can refer to a framework that coordinates the operation of protocols, connections, modules, and devices so that they can be executed appropriately and on schedule. In some embodiments, the operation may be performed using one or more logic elements, including a programmable logic controller (PLC), programmable logic array (PLA), programmable array logic (PAL), general-purpose logic array (GLA), composite programmable logic decision (CPLD), field-programmable gate array (FPGA), or application-specific integrated circuit (ASIC). The control system may comprise one or more network communication protocols, which may be standard network communication protocols, non-standard network communication protocols, or any combination thereof. In some embodiments, standard network communication protocols include Process Fieldbus (Profibus), Process Fieldnet (Profinet), Highway Addressable Remote Transducer (HART), Distributed Network Protocol (DNP3), Modbus, Open Platform Communications (OPC), Building Automation and Control Network (BACnet), Common Industrial Protocol (CIP), or Ethernet for Control Automation Technology (EtherCAT), PCI, NVMe, SAS, SATA, Fibre Channel, and USB. Data storage systems may include industrial, manufacturing, or processing facilities. Such facilities may support large-scale purposes such as the synthesis, storage, or retrieval of information stored in biomolecules. Data storage systems may include one or more of the following: PLCs, remote terminal units, intelligent electronic devices, engineering workstations, human-machine interfaces (HMIs), data historians, communication gateways, and front-end processors. In some embodiments, data storage systems may have controllable states distinct from process steps. In some embodiments, data storage systems may use open communication protocols.In some examples, the logic element comprises one or more of the following: motherboard 1 312, motherboard 2 314, FPGA 313, and control unit 315 (Figures 3B-3C).

[0036]

[0072] Logic elements can be used to control the operation of a data storage unit, including synthesis units, storage units, rack interfaces, or other components of a data storage unit. In some examples, logic elements are used to control the functionality of one or more flow cell blocks. In some examples, logic elements are used to control the functionality of one or more flow cells. In some examples, at least 2, 4, 6, 8, 10, 12, 16, or at least 24 flow cells are controlled per logic element. In some examples, approximately 2, 4, 6, 8, 10, 12, 16, or approximately 24 flow cells are controlled per logic element.

[0037]

[0073] A non-exclusive example of a system for data storage, along with a feedback loop, is shown in Figure 8. The feedback loop can generally include a user 1805 who can interact with the system via a controller 1835 (e.g., a PLC), for example, via a human-machine interface (HMI). The HMI may be a user interface (e.g., a GUI) that connects a person or user to one or more components of the system (e.g., equipment, a network, etc.). In some examples, the user may send input to the controller 1835, for example, as a query, regarding the state or function of the system components. In some examples, the query relates to all or part of an item of information stored in the system, such as the location, duration, or metadata of the information stored in the system.

[0038]

[0074] The controller 1835 may send output to the user 1805. The output may include responses to queries, which can be provided via the HMI and displayed on the user interface. In some examples, the interface includes S3 / OCI or cloud attachment. In some examples, the controller 1835 sends status information about the ICS components to the HMI, which is then provided to the user 1805. In some examples, the controller 1835 implements a control strategy using a system that includes a microprocessor for managing the components within the system.

[0039]

[0075] In some cases, the components may be physical devices such as equipment within the system. These physical devices may be used to store or retrieve information in biomolecules. For example, a physical device may be part of a synthesizer unit 1810, a storage unit 1815, an amplification unit 1820, or a sequencer unit 1825. In some examples, a physical device includes one or more components shown in Figures 2-6. In some examples, a physical device includes a robotic system 1830 that may be used to transport or handle biomolecules within the system. In some cases, the equipment may be on-site or remotely controlled. In some examples, a controller 1835 controls one or more physical devices within the system, such as control motors, valves, or switches.

[0040]

[0076] The controller 1835 can control a physical device based on one or more measurements obtained from sensors in the system. In some examples, the sensors are integrated into one or more modules (e.g., a synthesizer unit 1810, a storage unit 1815, an amplification unit 1820, or a sequencer unit 1825, a robotic system 1835, a flow cell, or a biomolecular extraction system). In some examples, the sensors determine when and how the physical device should operate. For example, the sensors may be built-in sensors as part of a control unit equipped with actuators. In some cases, the measurements may be physical measurements obtained from the sensors, such as pressure, volume, temperature, humidity, torque, vacuum, motion, flow rate (e.g., filling rate or discharge rate), or the orientation angle of the device (e.g., a flow cell). In some cases, the sensors are standalone sensors. In further examples, the controller 1835 receives commands from the user 1805, for example via an HMI, for physical devices to perform functions (e.g., pump operation, agitator operation, conveyor belt operation, etc.).

[0041]

[0077] sensor

[0078] The operation within the system or data from sensors may be supplied to one or more software modules for analyzing the data in the storage system, as described herein. For example, the data may be sensor data from one or more compartments or modules within the system, and an algorithm may be used to monitor one or more parameters. In some examples, the algorithm may be used to monitor patterns in the sensor data and detect anomalies, e.g., irregular sensor data from one or more compartments, and optionally alert the user through the HMI. In another example, the data may be information items or sequencing data, and an algorithm may be used to convert the data into another format (e.g., converting information items into nucleic acid sequences or vice versa). In some examples, the algorithm includes an error correction scheme that may be used to correct errors that occur during processing in the data storage system. In some examples, the sensor detects one or more of the following: pressure, temperature, humidity, salinity, light, UV, O2, flow rate, temperature gradient, electrical connection and status, and synthetic feedback data.

[0042]

[0079] Synthesis Unit

[0080] A system for data storage may include the synthesizer unit 1810 shown in Figure 8. The synthesizer unit can be used to synthesize biomolecules, such as multiple polynucleotides, that encode digital information. In some examples, the system comprises two or more synthesizer units 1810. Polynucleotides may be synthesized using the methods provided herein or any other suitable synthesis methods known in the art. Fluid and / or electronic control of polynucleotide synthesis in the synthesizer unit 1810 may be carried out by a controller 1835. In some examples, electronics within the synthesizer unit 1810 communicate with the controller 1835. In some examples, the synthesizer unit 1810 has an input for receiving a DNA sequence. In some examples, the synthesizer unit 1810 has an input for receiving fluid for polynucleotide synthesis. In some examples, the synthesizer unit 1810 has an output for eluting the synthesized polynucleotides. In some examples, the synthesized polynucleotides are transferred to another component of the system, such as a storage unit, amplification chamber, or sequencing unit, in non-limiting examples. In some examples, the synthesis unit includes one or more of the following: a computer controller (312-315), a flow cell block 311, an extraction stage 316, a storage unit 317, a robot interface, and a rack interface.

[0043]

[0081] reservoir

[0082] A data storage unit may include a reservoir or one or more reservoirs. In some examples, the reservoir is connected to or can be connected to a flow cell (via a flow cell block). In some examples, the flow cell is oriented at one or more angles (e.g., Figure 5C). In some examples, the flow cell is oriented such that α > 0°, β > 0°, or both. In some examples, the flow cell is oriented such that α ≤ 90°, β ≤ 45°, or both. In some examples, the flow cell is oriented such that 0° < α ≤ 90°, 0° < β ≤ 45°, or both. In some examples, the flow cell is oriented such that the plane of the cavity is substantially parallel to a bodily force such as gravity. In some examples, the reservoir and flow cell are stably associated using a base station, platform, or any other suitable equipment (e.g., implementation equipment such as a flow cell block). In some examples, the reservoir is part of a fluid distribution assembly that can be used to distribute a fluid (e.g., water, aqueous media, organic solvents, ionic liquids, etc.). In some examples, the system includes multiple reservoirs, each containing a different fluid. In some examples, the reservoirs contain liquids for extracting multiple biomolecules from a substrate. In some examples, the system may have multiple reservoirs, each containing a liquid (e.g., water, IPA, TBA (tert-butylamine), etc.) for biomolecule synthesis, storage, or acquisition. Those skilled in the art will understand that the size of the fluid-containing reservoirs can be easily adjusted. The size of the reservoirs may be, but are not limited to, about 10 mL, 25 mL, 50 mL, 75 mL, 100 mL, 250 mL, 500 mL, 750 mL, 1 L, 1.25 L, 1.5 L, 1.75 L, or about 2 L. In some examples, the reservoir size is approximately 10 mL to 2 L, 10 mL to 500 mL, 10 mL to 100 mL, 50 mL to 1 L, 50 mL to 500 mL, 50 mL to 100 mL, 100 mL to 2 L, 100 mL to 1 L, or 100 mL to 500 mL. In some examples, the reservoir is located inside the data storage unit or rack unit. In some examples, the reservoir is located outside the data storage unit or rack unit.In some examples, the size of the external reservoir is approximately 10L-2L, 10L-500L, 10L-100L, 50L-1L, 50L-500L, 50L-100L, 100L-2L, 100L-1L, or 100L-500L. In some examples, the reservoir is integrated into the storage unit.

[0044]

[0083] In some examples, the fluid distribution assembly includes pumps for moving fluid to or from one or more reservoirs. In some examples, the fluid distribution assembly includes a manifold 310, a valve assembly, or both. In certain embodiments, the assembly includes a mechanism for delivering a predetermined amount of fluid to a flow cell. The fluid is distributed, in some examples, by a pumping mechanism. Standard pumping techniques for pumping fluids known in the art may be employed in the system. Non-limiting examples of pumping include peristaltic pumps, pressurized fluidized beds, positive displacement pumps, and means such as syringe pumps. In some examples, the system additionally includes heating and / or cooling and / or insulating elements for controlling the temperature in various fluid reservoirs, in flow cells, or in mechanisms for transferring fluid between reservoirs and flow cells (e.g., manifolds), or in any combination thereof. In some examples, a valve bank 308 is used to control the movement of fluid. In some examples, a fluid manifold 310 is used to control the distribution and movement of fluid. In some examples, the manifold 310 has one or more valves.

[0045]

[0084] In some examples, the manifold 310 is connected (directly or indirectly) to or can be connected to one or more reservoirs. In this way, different fluid reagents can be brought into contact with the substrate within the flow cell. In some examples, reagents for carrying out various steps in the synthesis of biomolecules (e.g., nucleic acids or polypeptides) are introduced sequentially into the flow cell.

[0046]

[0085] The system may include an upper manifold and a bottom manifold. In some examples, the upper and bottom manifolds are connected to the same one or more reservoirs. In some examples, the upper and bottom manifolds are connected to one or more different reservoirs. In some examples, the reservoirs include a waste reservoir, a sample collection reservoir, or one or more reservoirs, each containing a different fluid. In some examples, both the upper and bottom manifolds are connected to a waste reservoir. In some examples, the upper manifold, the bottom manifold, or both have separate waste reservoirs. In some examples, the bottom manifold is connected to a sample collection reservoir. In some examples, the upper manifold is connected to a sample collection reservoir. In some examples, the bottom manifold is connected to one or more reservoirs, each containing a different fluid. In some examples, the upper manifold is connected to one or more reservoirs, each containing a different fluid. In some examples, the upper manifold is connected to, or can be connected to, a pump for moving fluid from the flow cell.

[0047]

[0086] Each manifold can be controlled independently. In some examples, each manifold includes multiple valves that can open and close paths between components of the system. In some examples, the fluid passing through the upper manifold, the bottom manifold, or both can be controlled independently, for example, by the use of valves that are operated automatically or manually. In some examples, the fluid passing through the system is controlled via a controller coupled to one or more actuators that open and close valves connected to a flow cell, reservoir, or pump. In such cases, the controller is used to control the flow rate or velocity of the fluid or gas throughout the system.

[0048]

[0087] In some examples, the system includes an upper manifold communicating with a portion of the flow cell having at least one upper opening. In some examples, the upper manifold includes a conduit communicating with the flow cell having at least one upper opening. In some examples, the upper manifold includes at least one opening that connects to or has the same spread as at least one upper opening of the flow cell. In some examples, the system includes a bottom manifold communicating with a portion of the flow cell having at least one bottom opening. In some examples, the bottom manifold includes a conduit communicating with the flow cell having at least one bottom opening. One or more components of the conduit or pipe connection system may be any suitable material known in the art, but not limited to PFA in some examples. In some examples, the conduit (or routing) may include, but is not limited to, diameters of about 1 / 4 inch, 1 / 8 inch, 1 / 16 inch, or 1 / 32 inch. In some examples, the bottom manifold includes at least one opening that connects to or has the same spread as at least one bottom opening of the flow cell. In some examples, when the flow cell is operating, the upper manifold is distal to the surface on which the portion of the flow cell containing at least one bottom opening is located. In some examples, the upper manifold can be used to introduce or fill a fully charged flow cell with fluid.

[0049]

[0088] In some examples, a system including a flow cell includes multiple upper and lower submanifolds, which allow fluids (liquid or gas) to enter the flow cell, respectively, through common upper and lower conduits. In some examples, the multiple upper and lower submanifolds are connected to one or more separate distribution lines. In yet another embodiment, the upper and lower submanifolds may be connected via a common distribution line, but the fluids passing through the upper or lower manifolds may be controlled independently by appropriately placed valves (e.g., via a controller).

[0050]

[0089] The system may further include a vacuum source. In some examples, the vacuum source is connected to or in communication with the flow cell. In some examples, the system includes one or more electronic sensors, mechanical sensors, or both for sensing the state of the flow cell. In some examples, the controller described herein is programmed to regulate the flow of fluid in the system through one or more sensors. In some examples, the system includes a fluid level sensor, one or more pressure transducers, one or more pressure regulators, manually or automatically operated valves and / or pumps.

[0051]

[0090] The system may further include mechanisms for facilitating the movement of the substrate into and out of the cavity of the flow cell, as described herein. In non-limiting examples, the system may include a lift mechanism for positioning the substrate into and / or lifting the substrate out of the cavity of the flow cell in a controlled manner, for example, manually or automatically.

[0052]

[0091] Flow cell block (FC block)

[0092] In some examples, the synthesis unit includes a flow cell block 311. In some examples, the flow cell block configuration is shown in Figures 4D-4E. In some examples, the flow cell block includes multiple flow cells 401. In some examples, the flow cells include one or more solid supports 402 for biomolecule synthesis (e.g., polynucleotides). In some examples, the flow cell block 311 includes inlets 404 and outlets 405 for reagents and waste, respectively. In some examples, the flow cell block includes inlets 406 and outlets 407 for gas (e.g., nitrogen or argon). In some examples, the ports (or inlets / outlets) are controlled by one or more valves. In some examples, the synthesis unit includes about 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 24, 32, 48, 64, 96, 128, 256, 512, or about 1024 flow cell blocks. In some examples, the synthesis unit includes at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 24, 32, 48, 64, 96, 128, 256, 512, or at least 1024 flow cell blocks. In some examples, the synthesis unit includes 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 24, 32, 48, 64, 96, 128, 256, 512, or 1024 or fewer flow cell blocks. In some examples, the synthesis unit includes 4-24, 4-20, 4-16, 4-12, 4-8, 6-24, 6-16, 6-12, 6-10, 8-24, 8-16, 8-12, 10-24, 12-24, or 16-24 flow cell blocks. In some examples, the flow cell block includes one or more flow cells comprising one or more solid supports configured for polynucleotide synthesis, and one or more flow cells comprising one or more ports for the exchange of gas, synthesis reagents and / or extracted polynucleotides. In some examples, the flow cell block includes a plurality of valves 403. In some examples, each valve controls the flow of fluid into each flow cell 401. In some examples, each flow cell 401 comprises at least one solid support 402. In some examples, each solid support 402 is double-sided or single-sided.A flow cell block may include different arrangements of flow cells. In some examples, the flow cells are arranged nearly perpendicularly within the flow cell block (e.g., α = 90°, as shown in Figure 5C). In some examples, the flow cells are arranged at an angle to the x-plane or y-plane (e.g., Figure 4E). In some examples, the flow cell block further includes a number of connectors and / or valves configured to distribute reagents or gases to individual flow cells. In some examples, the flow cell block includes inlets and outlets for each flow cell. In some examples, the flow cell block includes inlets and outlets for each flow cell for liquids. In some examples, the flow cell block includes inlets and outlets for each flow cell for gases (e.g., Figure 4D). In some examples, the flow cell block includes at least one manifold. In some examples, the manifold is shown in Figures 4B-4C. In some examples, the manifold includes a reagent distribution manifold. In some examples, at least one reagent distribution manifold is connected to one or more of the following: gas supply, flow waste, supply main waste, and supply inlet.

[0053]

[0093] In some examples, the flow cells 401 are spaced apart within the synthesis unit. In some examples, the flow cell blocks 311 include spacings of at least 10, 25, 50, 75, 100, 125, 150, 175, 200, or at least 250 mm. In some examples, the flow cell blocks include spacings of 10, 25, 50, 75, 100, 125, 150, 175, 200, or 250 mm or less. In some examples, the flow cell blocks include spacings of 10-300, 10-250, 10-225, 10-200, 10-175, 10-150, 10-125, 10-100, 10-75, 10-50, 25-300, 25-275, 25-250, 25-200, 25-175, 25-150, 25-125 This includes intervals of 25-100, 50-300, 50-250, 50-275, 50-225, 75-300, 75-250, 75-200, 75-150, 75-126, 75-175, 100-300, 100-250, 100-200, 150-300, 200-500, or 250-300 mm.

[0054]

[0094] Any combination of flow cell blocks 311 and any number of flow cells per block can be used in a synthesis unit. In some examples, the synthesis unit contains at least 12, 24, 36, 48, 64, 72, or 96 flow cells. In some examples, the synthesis unit contains about 12, 24, 36, 48, 64, 72, or about 96 flow cells. In some examples, the synthesis unit contains 12, 24, 36, 48, 64, 72 or fewer flow cells, or 96 or fewer flow cells. In some examples, the synthesis unit includes flow cells of 4-128, 4-96, 4-64, 8-128, 8-96, 8-96, 8-64, 16-256, 16-128, 16-96, 16-72, 16-64, 24-256, 24-128, 24-96, 48-256, 48-128, 64-256, 64-128, 64-96, 72-128, or 72-256.

[0055]

[0095] The flow cells 401 within the flow cell block 311 may have different pitches. In some examples, the flow cells have pitch distances of 1, 2, 5, 7, 8, 9, 10, 11, 12, 16, 20, or 25 mm or less. In some examples, the flow cells have pitch distances of at least 1, 2, 5, 7, 8, 9, 10, 11, 12, 16, 20, or at least 25 mm. In some examples, the flow cells have pitch distances of approximately 1, 2, 5, 7, 8, 9, 10, 11, 12, 16, 20, or approximately 25 mm. In some examples, the flow cells include pitch distances of 1-20, 1-15, 1-12, 1-11, 1-10, 1-9, 2-20, 2-15, 2-12, 2-10, 5-20, 5-15, 5-12, 5-10, 5-9, 7-15, 7-20, 7-25, 8-20, 8-15, 9-15, 9-20, 12-20, or 12-15 mm.

[0056]

[0096] Flow Cell

[0097] A flow cell 401, or a system or platform including a flow cell (e.g., having a valve 400), may be connected to or have the same extent as one or more components of a system such as a synthesizer unit 1810, a storage unit 1815, an amplification unit 1820, or a sequencer unit 1825. In some examples, the flow cell provided herein is used for biomolecule synthesis, biomolecule extraction, or both. In alternative examples, synthetic polynucleotides may be transferred to a flow cell or a system including a flow cell for the extraction of all or part of biomolecules from a synthetic surface. In some examples, once the polynucleotides have been cleaved from the surface, they are collected and, in non-limiting examples, transferred to another component of a system such as a storage unit 1815, an amplification unit 1820, or a sequencer unit 1825. In some examples, the flow cell is directed to maximize the recovery of liquid from the flow cell by adjusting an angle α or β as defined herein (e.g., Figure 5C). Furthermore, an apparatus comprising at least one logic element for performing one or more operations of the biomolecular synthesis and / or extraction platform provided herein may communicate with, or be part of, a controller 1835 of a larger data storage system. In some examples, the flow cell includes an input port and an output port. In some examples, the output port is configured to direct the liquid to a waste or extraction stage. In some examples, the flow cell is depicted as shown in Figure 2A. An exemplary schematic diagram showing the fluid routing of the flow cell block and individual flow cells is shown in Figure 6. In some examples, the flow cell includes an inlet and an outlet. In some examples, the flow cell inlet also includes a bypass for transfer to an extraction stage. In some examples, the flow cell includes an inlet for liquid reagents. In some examples, the flow cell includes an inlet for gas. In some examples, the gas outlet is connected to the flow cell outlet and the waste outlet. In some examples, fluid movement through the flow cell is achieved by positive pressure at the inlet. In some examples, fluid movement through the flow cell is achieved by vacuum at the outlet.

[0057]

[0098] In some examples, the flow cell 401 comprises a housing that defines a flow chamber, i.e., a cavity. In some examples, the flow cell includes an opening for receiving a substrate. The substrate may include biomolecules such as polynucleotides or proteins. In some examples, the cavity may contain the substrate and be fluidically sealed. In some examples, the opening of the flow cell may be configured to be sealable after the array substrate is placed therein, thereby preventing leakage of fluid from the flow cell through the opening. Such a seal may include a flexible material having sufficient flexibility or compressibility to form a fluid-sealed seal that can be maintained under the increased pressure encountered in the use of the device. Flexible members may be, for example, rubber, flexible plastics, flexible resins, and combinations thereof. In one embodiment, the flexible material is substantially inert to the fluid introduced into the device and does not interfere with the reactions that occur within the device. The flexible member may be a gasket and can be any shape, such as circular, elliptical, or rectangular. For example, in certain embodiments, the flexible member may be in the form of an O-ring (e.g., an FFKM Karles O-ring).

[0058]

[0099] In some examples, the flow cell 401 may include two halves. In some examples, the two halves can be placed in close proximity to enclose a substrate and fluidly seal the substrate. In some examples, if the flow chamber includes two halves, the halves may be stably connected by providing a fitting element (e.g., a projection on one half that fits into the opening of the other half). However, in another embodiment, the two halves may be stably connected by a clamp or other pressure sealing mechanism. In one embodiment, the two halves are sealable and engage during a reaction step (e.g., a synthesis step), and at other times severable so as to allow the support to be placed in and removed from the chamber of the flow cell. Movement of one half relative to the other can be achieved, for example, by a piston. Movement can be electronically controlled by means conventional in the art. In some examples, the two halves of the flow cell are mechanically joined. In some examples, the two halves of the flow cell are joined using one or more gaskets. In some examples, the two halves of the flow cell are joined using an adhesive. In some cases, the adhesive includes epoxy, polyurethane, or other adhesives.

[0059]

[0100] The dimensions of the flow cell 401, the cavity within it, or both can be varied. In some examples, the dimensions of the flow cell, the cavity, or both vary depending on the dimensions of the substrate placed within the flow cell. In some examples, the substrate includes an array on which the compound is synthesized. In some examples, the cavity has a first length and a second length (e.g., length and width). In some examples, the cavity is substantially rectangular. In some examples, the first length, the second length, or both of the cavity are approximately 15–22 mm. In some examples, the first length, second length, or both of the cavity are approximately 15-16, 15-17, 15-18, 15-19, 15-20, 15-21, 15-22, 16-17, 16-18, 16-19, 16-20, 16-21, 16-22, 17-18, 17-19, 17-20, 17-21, 17-22, 18-19, 18-20, 18-21, 18-22, 19-20, 19-21, 19-22, 20-21, 20-22, or 21-22 mm. In some examples, the first length, second length, or both of the cavity is approximately 15, 15.5, 16, 16.5, 17, 17.5, 17.9, 18, 18.5, 19, 19.5, 19.9, 20, 20.5, 21, 21.5, or 22 mm. In some examples, the first length, second length, or both of the cavity is at least approximately 15, 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, 20, 20.5, 21, or 21.5 mm. In some examples, the first, second, or both lengths of the cavity are approximately 15.5, 16, 16.5, 17, 17.5, 18, 18.5, 19, 19.5, 20, 20.5, 21, 21.5, or 22 mm. In some examples, the first length of the cavity is approximately 17.9 mm, and the second length is approximately 19.9 mm. In some examples, the cavity includes a third length (e.g., height). In some examples, the cavity is formed within a recess in the body of the flow cell. In some examples, the third length of the cavity is approximately 0.1–0.5 mm.In some examples, the third length of the cavity is approximately 0.1-0.15, 0.1-0.2, 0.1-0.25, 0.1-0.3, 0.1-0.35, 0.1-0.4, 0.1-0.45, 0.1-0.5, 0.15-0.2, 0.15-0.25, 0.15-0.3, 0.15-0.35, 0.15-0.4, 0.15-0.45, 0.15-0.5, 0.2-0.25, 0.2-0.3, 0. The lengths are 2-0.35, 0.2-0.4, 0.2-0.45, 0.2-0.5, 0.25-0.3, 0.25-0.35, 0.25-0.4, 0.25-0.45, 0.25-0.5, 0.3-0.35, 0.3-0.4, 0.3-0.45, 0.3-0.5, 0.35-0.4, 0.35-0.45, 0.35-0.5, 0.4-0.45, 0.4-0.5, or 0.45-0.5 mm. In some examples, the third length of the cavity is approximately 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, or 0.5 mm. In some examples, the third length of the cavity is at least approximately 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, or 0.45 mm. In some examples, the third length of the cavity is at most approximately 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, or 0.5 mm. In some examples, the cavity height is much smaller compared to the length and width, making the flow cell a Hele-Shaw flow cell. In some examples, the dimensions of the substrate are smaller than the dimensions of the cavity. In some examples, the dimensions of the flow cell are larger than the dimensions of the cavity.

[0060]

[0101] The volume that can be accommodated by the cavity of the flow cell 401 may vary. In some examples, the volume within the cavity ranges from approximately 22 to approximately 242 mm³. 3 In some cases, the volume inside the cavity is approximately 50 to 200 mm³. 3In some examples, the volume inside the cavity is approximately 22-45, 22-53, 22-68, 22-89, 22-95, 22-115, 22-125, 22-145, 22-194, 22-242, 45-53, 45-68, 45-89, 45-95, 45-115, 45-125, 45-145, 45-194, 45-242, 50-200, 53-68, 53-89, 53-95, 53-115, 53-125, 53-145, 53-194, 53-242, 68 ~89, 68~95, 68~115, 68~125, 68~145, 68~194, 68~242, 89~95, 89~115, 89~125, 89~145, 89~194, 89~242, 95~115, 95~125, 95~145, 95~194, 95~242, 115~125, 115~145, 115~194, 115~242, 125~145, 125~194, 125~242, 145~194, 145~242, or 194~242mm 3 In some cases, the volume of the cavity is approximately 22, 45, 50, 53, 68, 89, 95, 115, 125, 145, 194, 200, or 242 mm³. 3 In some cases, the volume of the cavity is at least approximately 22, 45, 50, 53, 68, 89, 95, 115, 125, 145, 194, or 200 mm³. 3 In some cases, the volume of the cavity is approximately 45, 50, 53, 68, 89, 95, 115, 125, 145, 194, 200, or 242 mm³ at its maximum. 3 That is the case.

[0061]

[0102] The flow cell device 401 provided herein may include an insulating material around at least a portion of the device. The flow cell device can expose a substrate within a cavity to a fluid flow, and a first end and a second end of the substrate are exposed to a fluid containing substantially the same composition at a given time interval. In some examples, the flow cell may be used to extract biomolecules from a substrate. In some examples, the flow cell may be used to carry out in situ synthesis of biomolecules (e.g., polynucleotides or polypeptides) on a substrate.

[0062]

[0103] The flow cell 401 may have openings (e.g., ports and / or valves). In some examples, the flow cell includes at least two openings. In some examples, the flow cell includes a first opening and a second opening, e.g., an inlet and an outlet. In some examples, during operation, the outlet port is located perpendicularly above the inlet port. In some examples, the flow cell includes 1 to 25 inlet holes, outlet holes, or both. In some examples, the flow cell has 1 to 2, 1 to 5, 1 to 8, 1 to 10, 1 to 12, 1 to 15, 1 to 20, 1 to 22, 1 to 25, 2 to 5, 2 to 8, 2 to 10, 2 to 12, 2 to 15, 2 to 20, 2 to 22, 2 to 25, 5 to 8, 5 to 10, 5 to 12, 5 to 15, 5 to 20, 5 to 22, 5 to 25, 8 to 10, The flow cell includes 8-12, 8-15, 8-20, 8-22, 8-25, 10-12, 10-15, 10-20, 10-22, 10-25, 12-15, 12-20, 12-22, 12-25, 15-20, 15-22, 15-25, 20-22, 20-25, or 22-25 inlet holes, outlet holes, or both. In some examples, the flow cell includes 1, 2, 5, 8, 10, 12, 15, 20, 22, or 25 inlet holes, outlet holes, or both. In some examples, the flow cell includes at least 1, 2, 5, 8, 10, 12, 15, 20, or 22 inlet holes, outlet holes, or both. In some examples, a flow cell contains up to 2, 5, 8, 10, 12, 15, 20, 22, or 25 inlet holes, outlet holes, or both. In some examples, the number and location of inlet holes, outlet holes, or both vary due to bias flow across different regions of the flow cell. However, in some examples, the number of inlet and outlet holes is uniform to provide unbiased flow. In some examples, the inlet holes are uniformly spaced along the bottom of the cavity. In some examples, the outlet holes are uniformly spaced along the top of the cavity. In some examples, the inlet holes are not uniformly spaced along the bottom of the cavity. In some examples, the outlet holes are not uniformly spaced along the top of the cavity. In some examples, uniform flow is achieved if the inlets and outlets are symmetrical. In some examples, uniform flow is achieved if the inlets and outlets are symmetrical, but the spacing between the inlet or outlet holes is not uniform.

[0063]

[0104] At least two openings in the cavity may vary in size. At least two openings may be perforated in the flow cell housing. In some examples, at least two openings are located on both sides of the cavity, for example, at the diagonal corners of a rectangular cavity, or along the opposite side of a rectangular cavity. In some examples, each of the at least two openings (e.g., an inlet hole and an outlet hole) is about 100 to about 500 μm. In some examples, each of at least two openings is approximately 100-150, 100-200, 100-250, 100-300, 100-350, 100-400, 100-450, 100-500, 150-200, 150-250, 150-300, 150-350, 150-400, 150-450, 150-500, 200-250, 200-300, The diameters are 200-350, 200-400, 200-450, 200-500, 250-300, 250-350, 250-400, 250-450, 250-500, 300-350, 300-400, 300-450, 300-500, 350-400, 350-450, 350-500, 400-450, 400-500, or 450-500 μm. In some examples, each of at least two openings has a diameter of approximately 100, 150, 200, 250, 300, 350, 400, 450, or 500 μm. In some examples, each of at least two openings has a diameter of at least about 100, 150, 200, 250, 300, 350, 400, or 450 μm. In some examples, each of at least two openings has a diameter of up to about 150, 200, 250, 300, 350, 400, 450, or 500 μm. In some examples, one or more openings in the cavity are connected to one or more manifolds for supplying fluid (e.g., liquid, gas, or both) into and / or out of the cavity. In some examples, the diameters of the first and second openings of at least two openings are the same. In some examples, the diameters of the first and second multiple openings of at least two openings are the same. In some examples, the diameters of the first and second openings of at least two openings are different.In some cases, the diameters of the first and second multiple openings among at least two openings are different.

[0064]

[0105] In some examples, high pressure at one or more openings (e.g., inlet holes) equalizes the pressure within the manifold. In some examples, the manifold provides a mechanism for removing bubbles or reducing the presence of bubbles in one or more openings (e.g., inlet holes). In some examples, the diameter of the outlet hole in a flow cell connected to or connectable to the manifold is larger than the diameter of the inlet hole, for example, at least about 1x larger, at least about 1.5x larger, at least about 2x larger, or at least about 4x larger. In some examples, the flow through the inlet and / or outlet holes to the manifold is controlled by providing valves whose opening and closing are controlled by a controller such as a microprocessor.

[0065]

[0106] In some examples, the roles of one or more manifolds can be reversed between or during the operation of the flow cell device. In some examples, it is advantageous to introduce fresh reagent from the top of the flow cell (for example, when the fresh reagent is less dense than the residual liquid). In some examples, a valve can be closed to increase the pressure in the manifold (e.g., the top manifold) to introduce liquid into the flow cell through the outlet, and a manifold (e.g., the bottom manifold) can be used to ventilate the flow cell device.

[0066]

[0107] The flow cell 401 may be positioned vertically using a stand or base. In some examples, the flow cell may include, or be connectable to, a base station or platform to which one or more fluid distribution stations can be stably associated (e.g., by mounts). In some examples, the flow cell is positioned on a stand or base using mounts so that the flow cell can be oriented at one or more angles.

[0067]

[0108] The flow cell 401 may be positioned vertically using a stand or base and / or rotated. In some examples, the flow cell may include a base station or platform to which one or more fluid distribution stations can be stably associated (e.g., by mounts), or may be connectable to them. In some examples, mounts are used to position the flow cell on a stand or base. In some examples, the stand or base allows the flow cell to be oriented at one or more angles, such as angles α or β as described herein. In some examples, a user or an automated system (e.g., a robotic system) is used to adjust or oriented the flow cell at one or more angles (e.g., angles α or β). In some examples, a flow cell oriented at angles α=90° and β=45° (e.g., Figure 5C) may include two or fewer openings. The cavity may be substantially rectangular or square. The fluid boundary of the cavity may be defined by an O-ring that can fluidly seal the cavity. In some examples, the flow cell includes a first opening at the bottom and a second opening at the top. In exemplary embodiments, when the cavity is filled, liquid can enter through the first opening (bottom) and any excess liquid can be removed through the second opening (top) (see, for example, “Filling” or “Washing” in Figure 5A, or “Filling” in Figure 5B). Furthermore, when the cavity is evacuated, air can be filled through the second opening (top) and the liquid in the cavity can be discharged through the first opening (bottom) (see, for example, “Drying” in Figure 5A or “Extraction” in Figure 5B).

[0068]

[0109] The flow cells provided herein can be integrated into systems such as systems for synthesizing and / or extracting materials from a surface (e.g., Figures 4A–4E). In some examples, the system extracts biomolecules, such as polynucleotides, from a substrate. In some examples, a flow cell containing a cavity is oriented such that the plane of the cavity is substantially parallel to a physical force in the system, such as gravity. In some examples, a substrate containing multiple biomolecules is placed within the cavity of the substrate and fluidly sealed (e.g., using O-rings). The flow cell may include a heating or cooling unit configured to vary the temperature of the flow cell. In some examples, the flow cell includes a heating unit. In some examples, the flow cell includes a cooling unit. In some examples, the flow cell includes a controller that adjusts the flow cell temperature. In some examples, the flow cell is heated during a deprotection or extraction step. In some examples, the heating or cooling unit is configured to adjust the temperatures of multiple flow cells, such as a flow cell block.

[0069]

[0110] solid support

[0111] A synthesizer unit may include one or more solid supports. In some examples, a solid support 402 is shown in Figure 4E. In some examples, the solid support includes a chip. The solid support may include a surface for polynucleotide synthesis. In some examples, the flow cell includes multiple solid supports. In some examples, the solid supports are in fluid communication with the flow cell. In some examples, the solid support includes multiple loci for the synthesis of biomolecules. In some examples, these loci are addressable. In some examples, different biomolecules are synthesized at each addressable locus. In some examples, the biomolecules include polynucleotides. In some examples, polynucleotides with different sequences are synthesized at each addressable locus. In some examples, control over individual loci includes masking. In some examples, masking includes control of reagent contact with one or more loci on the surface. In some examples, the reagents include deprotection reagents, coupling reagents, or other reagents used for polynucleotide synthesis. In some examples, a CMOS device is used to control synthesis at each locus on the surface. In some examples, the use of masking at each locus allows for the washing of reagents (coupling reagents, bulk reagents, washing solvents, or other reagents) across the entire surface. In some examples, masking controls which loci react with a particular reagent. In some examples, biomolecules are synthesized on both sides of a solid support.

[0070]

[0112] Different arrangements of solid supports may be used within the flow cell. In some examples, the flow cell contains multiple solid supports. In some examples, the flow cell contains at least 10, 20, 30, 50, 70, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, or at least 500 solid supports for polynucleotide synthesis. In some examples, synthesis is performed on two surfaces of the solid support (e.g., front and back). In some examples, the flow cell contains at least 10, 20, 30, 50, 70, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, or at least 500 surfaces for polynucleotide synthesis. In some examples, the flow cell contains approximately 10, 20, 30, 50, 70, 100, 120, 140, 160, 180, 200, 220, 240, 260, 280, 300, or approximately 500 surfaces for polynucleotide synthesis. In some examples, the structure contains 10-500, 25-500, 50-500, 100-500, 150-500, 100-300, 150-250, 200-300, 200-500, or 300-500 surfaces for polynucleotide synthesis. In some examples, the flow cell comprises widths of 5-10, 5-15, 5-20, 5-25, 10-25, 10-50, or 20-50 solid supports, and heights of 5-10, 5-15, 5-20, 5-25, 10-25, 10-50, or 20-50 solid supports.

[0071]

[0113] Solid supports can be constructed from a variety of materials. In some examples, the solid support, surface, or both include materials described herein. In some examples, the material includes metals or organic polymers. In some examples, the material includes steel (e.g., stainless steel) or other metal alloys. In some examples, the material includes polyethylene, polypropylene, or other polymers. In some examples, the structure includes flexible materials such as those provided herein. Exemplary flexible materials include, without limitation, modified nylon, unmodified nylon, nitrocellulose, and polypropylene. In some examples, the material includes rigid materials such as those provided herein. Examples of rigid materials include, without limitation, glass, quartz glass, silicon, silicon dioxide, silicon nitride, plastics (e.g., polytetrafluoroethylene, polypropylene, polystyrene, polycarbonate, and blends thereof), and metals (e.g., steel, gold, platinum). In some examples, the materials disclosed herein may be made of materials comprising silicon, polystyrene, agarose, dextran, cellulosic polymers, polyacrylamide, polydimethylsiloxane (PDMS), glass, or any combination thereof. In some examples, the materials disclosed herein are manufactured from a combination of the materials listed herein or any other suitable materials known in the art. In some examples, a storage chamber includes the materials described herein for a solid support.

[0072]

[0114] In some examples, the solid support has various dimensions. In some examples, the size of the solid support is about 40 - 120 mm × about 25 - 100 mm. In some examples, the size of the solid support is about 80 mm × about 50 mm. In some examples, the width of the solid support is at least, or about 10 mm, 20 mm, 40 mm, 60 mm, 80 mm, 100 mm, 150 mm, 200 mm, 300 mm, 400 mm, 500 mm, or more than 500 mm. In some examples, the height of the solid support is at least, or about 10 mm, 20 mm, 40 mm, 60 mm, 80 mm, 100 mm, 150 mm, 200 mm, 300 mm, 400 mm, 500 mm, or more than 500 mm. In some examples, the solid support is at least or about 100 mm 2 , 200 mm 2 , 500 mm 2 , 1,000 mm 2 , 2,000 mm 2 , 4,500 mm<00​​​​​​​​​​​​​​​​​​​​​In some examples, two or more solid supports are assembled. In some examples, the solid supports are interfaced together onto a larger unit. The interface connection may include the exchange of fluids, electrical signals, or other exchange media between the solid supports. This unit can interface with any number of servers, computers, or network devices. For example, multiple solid supports are incorporated into a rack unit that can be conveniently inserted into or removed from a server rack. The rack unit may contain any number of solid supports. In some examples, the rack unit contains at least 1, 2, 5, 10, 20, 50, 100, 200, 500, 1000, 2000, 5000, 10,000, 20,000, 50,000, 100,000, or more than 100,000 solid supports. In some examples, two or more solid supports are not interfaced with each other. Polynucleotides present on the solid supports (and the information stored thereon) can be accessed from the rack unit. Access includes the extraction of polynucleotides from a solid support, direct analysis of polynucleotides on a solid support, or any other method that enables the manipulation or identification of information stored in the polynucleotides. In some examples, the information is accessed from multiple racks, a single rack, a single solid support on a rack, a portion of a solid support, or a single locus on a solid support. In various examples, access involves interfacering the polynucleotides with additional equipment, such as a mass spectrometer, HPLC, sequencing instrument, PCR thermocycler, or other equipment that manipulates polynucleotides. In some examples, access to nucleic acid information is achieved by cleaving the polynucleotides from all or part of a solid support. Cleavage, in some examples, involves exposure to chemical reagents (ammonia or other reagents), electrical potential, radiation, heating, light, sound, or other forms of energy capable of manipulating chemical bonds. In some examples, cleavage occurs by placing the solid support in a flow cell.In some examples, the orientation of one or more flow cells is optimized to maximize the amount of liquid containing polynucleotides that can be recovered from the flow cell (e.g., angle α or β). In some examples, cleavage occurs by charging one or more electrodes near the polynucleotides. In some examples, electromagnetic radiation in the form of ultraviolet light is used to cleave the polynucleotides. In some examples, a lamp is used to cleave the polynucleotides, with a mask interposed at the location of ultraviolet light exposure to the surface. In some examples, a laser is used to cleave the polynucleotides, with ultraviolet light exposure to the surface controlled by the opening and closing of a shutter. In some examples, access to nucleic acid information (including removal / addition of racks, solid supports, reagents, polynucleotides, or other components) is fully automated.

[0074]

[0116] Solid supports as described herein include an active region. In some examples, the active region includes a region, cell, feature, or locus for nucleic acid synthesis. In some examples, the active region includes a region or locus for nucleic acid storage. In some examples, the region or locus includes one or more modules. In some examples, the region or locus includes one or more second modules. In some examples, the region is addressable. In some examples, the region is addressable through an electrode.

[0075]

[0117] The active area includes a variety of different dimensions. For example, the dimensions of the active area are approximately 1 mm to 50 mm x 1 mm to 50 mm. In some examples, the active area includes a width of at least or greater than approximately 0.5, 1, 1.5, 2, 2.5, 3, 5, 5, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, or 80 mm. In some examples, the active area includes a height of at least or greater than approximately 0.5, 1, 1.5, 2, 2.5, 3, 5, 5, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, or 80 mm.

[0076]

[0118] In some examples, the solid support has a large number of sites (e.g., spots) or positions for polynucleotide synthesis. In some examples, the solid support can be used for storing polynucleotides. In some examples, the solid support contains up to or about 10,000 × 10,000 positions in a given region. In some examples, the solid support contains about 1,000 to 20,000 × about 1,000 to 20,000 positions in a given region. In some examples, the solid support contains at least or about 10, 30, 50, 75, 100, 200, 300, 400, 500, 1,000, 2,000, 3,000, 4,000, 5,000, 6,000, 7,000, 8,000, 9,000, 10,000, 12,000, 14,000, 16,000, 18,000, 20,000 positions in a given region. The 00 positions include at least or approximately 10, 30, 50, 75, 100, 200, 300, 400, 500, 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, 10,000, 12,000, 14,000, 16,000, 18,000, and 20,000 positions. In some examples, the area is up to 0.25, 0.5, 0.75, 1.0, 1.25, 1.5, or 2.0 square inches. In some examples, the solid support contains loci with pitches of at least 0.1, 0.2, 0.25, 0.3, 0.4, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5, 6, 7, 8, 9, 10, or more than 10 μm. In some examples, the solid support contains loci with a pitch of approximately 5 μm. In some examples, the solid support contains loci with a pitch of approximately 2 μm. In some examples, the solid support contains loci with a pitch of approximately 1 μm. In some examples, the solid support contains loci with a pitch of approximately 0.2 μm. In some examples, the solid support contains loci with pitches of approximately 0.2 μm to 10 μm, 0.2 μm to 8 μm, 0.5 μm to 10 μm, 1 μm to 10 μm, 2 μm to 8 μm, 3 μm to 5 μm, 1 μm to 3 μm, or 0.5 μm to 3 μm. In some examples, the solid support contains loci with pitches of approximately 0.1 μm to 3 μm.

[0077]

[0119] In some examples, solid supports can be used for polynucleotide storage. In some examples, solid supports have large capacities for data storage. For example, the capacity of a solid support may be at least or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000 petabytes, or more than 1000 petabytes. In some examples, the capacity of a solid support is about 1 to about 10 petabytes or about 1 to about 100 petabytes. In some examples, the capacity of a solid support is about 100 petabytes. In some examples, polynucleotides are stored on a solid support for a certain period of time and subsequently extracted from the solid support using systems and methods provided herein. For example, polynucleotides on a solid support may be stored for days, months, years, or decades for the recovery or quality control of all or part of the polynucleotide information and subsequently extracted from the solid support using a flow cell. In some examples, one or more orientations of the flow cell are optimized to maximize the amount of liquid containing polynucleotides that can be recovered from the flow cell (e.g., angle α or β).

[0078]

[0120] In some examples, data is stored as an array of packets as droplets. In some examples, the array of packets is an addressable packet. In some examples, packets are addressable using electrodes. In some examples, data is stored as an array of packets as droplets on a spot. In some examples, data is stored as an array of packets as a drywell. In some examples, the array contains at least or approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 200 gigabytes of data, or more than 200 gigabytes. In some examples, the array contains at least or approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 200, or more than 200 terabytes of data. In some examples, the array contains at least or approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 200, or more than 200 petabytes of data. In some examples, the array contains at least or more than approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 200, or 200 exabytes of data. In some examples, information items are stored in the background of the data. For example, an information item encodes approximately 10 to approximately 100 terabytes of data and is stored in 1 petabyte of background data. In some examples, an information item encodes approximately 10 to approximately 100 petabytes of data and is stored in 1 petabyte of background data. In some examples, an information item encodes approximately 1 to approximately 100 exabytes of data and is stored in 1 petabyte of background data. In some examples, information items encode at least 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 300, 400, 500 terabytes, or more than 500 terabytes of data, and are stored in background data exceeding 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 300, 400, 500 petabytes, or more than 500 petabytes.In some examples, information items encode at least 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 300, 400, 500 petabytes, or more than 500 petabytes of data, and store in background data of 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 300, 400, 500 exabytes, or more than 500 exabytes. In some examples, plates contain multiple wells, e.g., 96, 384, 1024, or more wells.

[0079]

[0121] A data storage system including a solid support is provided herein, and after synthesis, the polynucleotides are collected in packets as one or more droplets. In some examples, the polynucleotides are collected in packets as one or more droplets and stored. In some examples, the number of droplets is at least or about 1, 10, 20, 50, 100, 200, 300, 500, 1000, 2500, 5000, 75000, 10,000, 25,000, 50,000, 75,000, 100,000, 1,000,000, 5,000,000, 10,000,000, 25,000,000, 50,000,000, 75,000,000, 100,000,000, 250,000,000, 500,000,000, or more than 750,000,000 droplets. In some examples, droplet volumes include diameters of 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, or greater than 100 μm (micrometers). In some examples, droplet volumes include diameters of 1–100 μm, 10–90 μm, 20–80 μm, 30–70 μm, or 40–50 μm.

[0080]

[0122] In some examples, the polynucleotides collected in the packets contain similar sequences. In some examples, the polynucleotides further contain non-identical sequences used as tags or barcodes. For example, non-identical sequences are used to index polynucleotides stored on a solid support and to later retrieve specific polynucleotides based on the non-identical sequences. Exemplary tag or barcode lengths include, but are not limited to, barcode sequences containing approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 or more base pairs. In some examples, the tags or barcodes contain lengths of at least or approximately 10, 50, 75, 100, 200, 300, 400, or more than 400 base pairs.

[0081]

[0123] A data storage system including a solid support is provided herein, in which polynucleotides are collected in packets containing redundancy. For example, a packet contains about 100 to about 1000 copies of each polynucleotide. In some examples, a packet contains at least or about 50, 75, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1200, 1400, 1600, 1800, 2000, or more than 2000 copies of each polynucleotide. In some examples, a packet has about 1000 to about 5000 times the synthetic redundancy. In some examples, the redundancy of synthesis is at least or about 500 times, 1000 times, 1500 times, 2000 times, 2500 times, 3000 times, 3500 times, 4000 times, 5000 times, 6000 times, 7000 times, 8000 times, or more than 8000 times. Polynucleotides synthesized using the solid support-based methods described herein include a variety of lengths. In some examples, the polynucleotides are synthesized and further stored on the solid support. In some examples, the polynucleotide length is about 100 to about 1000 bases. In some examples, polynucleotides contain at least or about 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, or more than 2000 base lengths.

[0082]

[0124] De novo polynucleotide synthesis

[0125] Provided herein are systems and methods for synthesizing libraries of polynucleotides on a substrate. In some examples, a library containing multiple polynucleotides from a coding scheme is synthesized. In some examples, a library containing multiple polynucleotides from a coding scheme encodes a pool of multiple pools. In some examples, a library containing multiple polynucleotides from a coding scheme encodes an index pool. In some examples, the method includes the use of electrochemical deprotection. In some examples, the substrate is a flexible substrate. In some examples, at least 10 10 , 10 11 , 10 12 , 10 13 , 10 14 , or 10 15 10 × 10¹⁶ bases are synthesized in one day. In some examples, at least 10 × 10¹⁶ bases are synthesized. 8 , 10×10 9 , 10×10 10 , 10×10 11 , or 10 x 10 12Nine polynucleotides are synthesized in one day. In some cases, each polynucleotide synthesized contains at least 20, 50, 100, 200, 300, 400, or 500 nucleic acid bases. In some cases, these bases are synthesized with an overall average error rate of less than approximately 1 per 100;200;300;400;500;1000;2000;5000;10000;15000;20000 bases. In some examples, such error rates are for at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, 99.5%, or more of the polynucleotides synthesized. In some examples, at least 90%, 95%, 98%, 99%, 99.5%, or more of the polynucleotides synthesized are identical to the predetermined sequence it encodes. In some examples, the error rates of polynucleotides synthesized on a substrate using the methods and systems described herein are less than about 1 / 200, less than about 1 / 1,000, less than about 1 / 2,000, less than about 1 / 3,000, or less than about 1 / 5,000. Individual types of error rates include mismatches, deletions, insertions, and / or substitutions of polynucleotides synthesized on the substrate. The term “error rate” refers to a comparison between the amount of synthesized polynucleotides as a population and the total amount of a predetermined polynucleotide sequence. In some examples, the synthesized polynucleotides disclosed herein contain 12 to 25 bases in tethering. In some examples, the tethering includes 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 bases or more.

[0083]

[0126] This specification describes methods, systems, apparatus, and compositions in which chemical reactions used in polynucleotide synthesis are controlled by electrochemistry. In some examples, the electrochemical reactions are controlled by any energy source, such as light, heat, radiation, or electricity. For example, electrodes are used to control the chemical reactions as all or part of individual gene loci on a surface. In some examples, electrodes are charged by applying an electric potential to them, thereby controlling one or more chemical steps in polynucleotide synthesis. In some examples, these electrodes are addressable. Any number of chemical steps described herein are controlled by one or more electrodes in some examples. Electrochemical reactions may include oxidation, reduction, acid / base chemistry, or other reactions controlled by electrodes. In some examples, electrodes produce electrons or protons used as reagents for chemical conversion. In some examples, electrodes directly produce reagents such as acids. In some examples, the acid is a proton. In some examples, electrodes directly produce reagents such as bases. Acids or bases are often used to cleave protecting groups or to influence the kinetics of various polynucleotide synthesis reactions, for example, by adjusting the pH of the reaction mixture. Electrochemically controlled polynucleotide synthesis reactions, in some examples, involve redox-active metals or other redox-active organic materials. In some examples, metal or organocatalysts are employed in these electrochemical reactions. In some examples, acids are produced from the oxidation of quinones.

[0084]

[0127] Control of chemical reactions is not limited to the electrochemical generation of reagents. Chemical reactivity can be indirectly influenced through biophysical changes in the substrate or reagent via an electric field (or gradient) generated by electrodes. In some examples, the substrate may be, but is not limited to, polynucleotides. In some examples, an electric field is generated that repels or attracts a particular reagent or substrate toward or from an electrode or surface. Such an electric field is generated in some examples by applying a potential to one or more electrodes. For example, a negatively charged polynucleotide is repelled away from a negatively charged electrode surface. Such repulsive or attractive forces of a polynucleotide or other reagent caused by a local electric field result in, in some examples, the movement of the polynucleotide or other reagent within or outside the region of the synthesis apparatus or structure. In some examples, electrodes generate an electric field that repels the polynucleotide away from the synthesis surface, structure, or apparatus. In some examples, electrodes generate an electric field that attracts the polynucleotide toward the synthesis surface, structure, or apparatus. In some examples, protons are repelled away from a positively charged surface, thus limiting contact between protons and the substrate or a portion thereof. In some examples, repulsive or attractive forces are used to allow or prevent reagents or substrates from entering specific areas of the synthesis surface. In some examples, contact between nucleoside monomers and polynucleotide chains is prevented by applying an electric field near one or both components. Such configurations allow for gating of specific reagents, potentially eliminating the need for protecting groups when the concentrations of reagents and / or substrates or the contact rate between them are controlled. In some examples, unprotected nucleoside monomers are used for polynucleotide synthesis. Alternatively, applying an electric field near one or both components promotes contact between nucleoside monomers and polynucleotide chains. In addition, applying an electric field to a substrate can alter its reactivity or conformation. In exemplary applications, the electric field generated by electrodes is used to prevent interaction with polynucleotides at adjacent loci. In some examples, the substrate is a polynucleotide optionally attached to the surface. When an electric field is applied, in some examples, the three-dimensional structure of the polynucleotide changes.Such alterations include folding or unfolding various structures, such as helices, hairpins, loops, or other three-dimensional nucleic acid structures. These alterations are useful for manipulating polynucleotides located within wells, channels, or other structures. In some examples, an electric field is applied to the nucleic acid substrate to prevent secondary structures. In some examples, the electric field eliminates the need for linker or solid support attachment during polynucleotide synthesis.

[0085]

[0128] A preferred method for polynucleotide synthesis on the substrates of this disclosure is the synthesis of a phosphoramidite base of DNA. In some cases, the reagents for the synthesis of the phosphoramidite base include one or a combination of a nucleoside phosphoramidite, an oxidizing agent, an activating agent, or a deblocking agent, or the solvent includes acetonitrile. In some examples, the method for synthesizing the phosphoramidite base includes the controlled addition of a phosphoramidite component, i.e., a nucleoside phosphoramidite, to a growing polynucleotide chain in a coupling step that forms a phosphytotryster bond between the phosphoramidite component and a nucleoside bound to the substrate. In some examples, the nucleoside phosphoramidite is provided to an activated substrate. In some examples, the nucleoside phosphoramidite is provided to the substrate together with an activating factor. In some examples, nucleoside phosphoramidites are provided to the substrate in an excess of 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100 times or more of the nucleoside bound to the substrate. In some examples, the addition of nucleoside phosphoramidites is carried out in an anhydrous environment, for example, in anhydrous acetonitrile. After the addition and binding of nucleoside phosphoramidites in the coupling step, the substrate is optionally washed. In some examples, the coupling step is repeated one or more times, with optionally a washing step interspersed between additions of nucleoside phosphoramidites to the substrate. In some examples, the polynucleotide synthesis method used herein includes one, two, three or more sequential coupling steps. In many cases, before coupling, the nucleoside bonded to the substrate is deprotected by removing a protecting group, which prevents polymerization. The protecting group can be any chemical group that prevents the elongation of the polynucleotide chain. In some cases, the protecting group is cleaved (or removed) in the presence of an acid. In some cases, the protecting group is cleaved in the presence of a base. In some cases, the protecting group is removed by electromagnetic radiation such as light, heat, or other energy sources. In some cases, the protecting group is removed through oxidation or reduction reactions.In some cases, the protecting group includes a triarylmethyl group. In some cases, the protecting group includes an aryl ether. In some cases, the protection includes a disulfide. In some cases, the protecting group includes an acid-unstable silane. In some cases, the protecting group includes an acetal. In some cases, the protecting group includes a ketal. In some cases, the protecting group includes an enol ether. In some cases, the protecting group includes a methoxybenzyl group. In some cases, the protecting group includes an azide. In some cases, the protecting group is 4,4'-dimethoxytrityl (DMT). In some cases, the protecting group is tert-butyl carbonate. In some cases, the protecting group is tert-butyl ester. In some cases, the protecting group includes a base-unstable group.

[0086]

[0129] Phosphoramidite polynucleotide synthesis methods optionally include a capping step after coupling. In the capping step, the growing polynucleotide is treated with a capping agent. The capping step generally prevents further chain elongation by unreacted 5'-OH groups bound to the substrate after coupling, thus preventing the formation of polynucleotides with internal base deletions. Furthermore, activation of phosphoramidites with 1H-tetrazole often results in a small reaction with the O6 position of guanosine. Although not theoretically constrained, during oxidation with I2 / water, this byproduct undergoes depurination, probably via O6-N7 migration. The purine-free site may be cleaved during the final deprotection of the polynucleotide, potentially reducing the yield of the full-length product. Modification at the O6 position can be removed by treatment with a capping reagent before oxidation with I2 / water. In some cases, including a capping step in polynucleotide synthesis reduces the error rate compared to synthesis without capping. For example, the capping step involves treating the polynucleotide bound to the substrate with a mixture of acetic anhydride and 1-methylimidazole. After the capping step, the substrate is optionally washed.

[0087]

[0130] Following the addition of the nucleoside phosphoramidite, and optionally after capping and one or more washing steps, the substrates described herein contain a bonded growing nucleic acid, which may be oxidized. The oxidation step involves oxidizing the phosphite triester to a tetracoordinate phosphate triester, which is a protected precursor of the naturally occurring phosphate diester nucleoside bond. In some examples, the phosphite triester is oxidized electrochemically. In some examples, the oxidation of the growing polynucleotide is achieved by treatment with iodine and water in the presence of a weak base such as pyridine, lutidine, or colidine, optionally. Oxidation may sometimes be carried out under anhydrous conditions using tert-butyl hydroperoxide or (1S)-(+)-(10-camphorsulfonyl)-oxaziridine (CSO). In some methods, a capping step is performed after oxidation. A second capping step allows for drying of the substrate, as residual water from oxidation can interfere with subsequent coupling. After oxidation, the substrate and the growing polynucleotides are optionally washed. In some examples, instead of this oxidation step, a sulfidation step is performed to obtain polynucleotide phosphorothioates, where an optional capping step can be carried out after sulfidation. Many reagents are capable of efficiently transferring sulfur, but are not limited to: 3-(dimethylaminomethylidene)amino)-3H-1,2,4-dithiazole-3-thione, DDTT, 3H-1,2-benzodithiol-3-one 1,1-dioxide, also known as Beaucage reagent, and N,N,N'N'-tetraethylthiuram disulfide (TETD).

[0088]

[0131] To carry out the subsequent nucleoside incorporation cycle via coupling, the protected 5' end (or 3' end, if synthesis proceeds from 5' to 3') of the growing polynucleotide bound to the substrate is removed, thus allowing the primary hydroxyl group to react with the adjacent nucleoside phosphoramidite. In some examples, the protecting group is DMT, and deblocking is carried out with trichloroacetic acid in dichloromethane. In some examples, the protecting group is DMT, and deblocking is carried out with electrochemically generated protons. Prolonged detritylation or detritylation with an acid stronger than the recommended solution can lead to increased depurination of the polynucleotide bound to the solid support, and consequently, a decrease in the yield of the desired full-length product. The methods and compositions described herein provide controlled deblocking conditions and limit undesirable depurination reactions. In some examples, the polynucleotide bound to the substrate is washed after deblocking. In some cases, efficient washing after deblocking contributes to a low error rate of the synthesized polynucleotide.

[0089]

[0132] Methods for synthesizing polynucleotides on substrates described herein may involve a series of steps: applying a protected monomer to the surface of a substrate feature to link it with the surface, a linker, or a pre-deprotected monomer; deprotecting the applied monomer to allow it to react with a subsequently applied protected monomer; and repeating the process of adding another protected monomer for linkage. One or more intermediate steps include oxidation and / or sulfurization. In some examples, one or more washing steps are performed before or after one or all of these steps.

[0090]

[0133] Methods for synthesizing polynucleotides on substrates described herein may include oxidation steps. For example, a method may involve the following series of steps: applying a protected monomer to the surface of a substrate feature to link it with the surface, a linker, or a pre-deprotected monomer; deprotecting the applied monomer to allow it to react with a subsequently applied protected monomer; adding another protected monomer for linking; and repeated oxidation and / or sulfidation. In some examples, one or more washing steps are performed before or after one or all of these steps.

[0091]

[0134] Methods for synthesizing polynucleotides on substrates described herein may further include the following steps: applying a protected monomer to the surface of a substrate feature to link it to either the surface, a linker, or a pre-deprotected monomer; deprotecting the applied monomer to allow it to react with a subsequently applied protected monomer; and repeated oxidation and / or sulfidation. In some examples, one or more washing steps are performed before or after one or all of these steps.

[0092]

[0135] Methods for synthesizing polynucleotides on substrates described herein may further include the following series of steps: applying a protected monomer to the surface of a substrate feature and linking it to either the surface, a linker, or a pre-deprotected monomer; and repeated oxidation and / or sulfidation. In some examples, one or more washing steps are performed before or after one or all of these steps.

[0093]

[0136] Methods for synthesizing polynucleotides on substrates described herein may further include the following steps: applying a protected monomer to the surface of a substrate feature to link it to either the surface, a linker, or a pre-deprotected monomer; deprotecting the applied monomer to allow it to react with a subsequently applied protected monomer; and repeated oxidation and / or sulfidation. In some examples, one or more washing steps are performed before or after one or all of these steps.

[0094]

[0137] In some examples, polynucleotides are synthesized with photo-dissociable protecting groups, where hydroxyl groups generated on the surface are blocked by the photo-dissociable protecting groups. When the surface is exposed to ultraviolet light, such as through a photomask, a pattern of free hydroxyl groups can be generated on the surface. These hydroxyl groups can react with photoprotected nucleoside phosphoramidites according to phosphoramidite chemistry. A second photomask can be added to expose the surface to ultraviolet light to generate a second pattern of hydroxyl groups, which can then be coupled with 5'-photoprotected nucleoside phosphoramidites. Similarly, patterns can be generated and oligomeric chains can be extended. Although not constrained by theory, the dissociability of photocleavable groups depends on the wavelength and polarity of the solvent used, and the photocleavage rate can be affected by the exposure time and light intensity. This method can leverage several factors, including the precision of mask alignment, the efficiency of photoprotection group removal, and the yield of the phosphoramidite coupling step. Furthermore, it can minimize unintended light leakage to adjacent sites. The density of synthesized oligomers per spot can be monitored by adjusting the load of leader nucleosides onto the synthesis surface.

[0095]

[0138] The surface of the substrates described herein, which provide support for polynucleotide synthesis, may be chemically modified to allow the cleavage of the synthesized polynucleotide chain from the surface. In some examples, the polynucleotide chain is cleaved at the same time as the deprotection of the polynucleotide. In some cases, the polynucleotide chain is cleaved after the deprotection of the polynucleotide. In an exemplary scheme, a trialkoxysilylamine such as (CH3CH2O)3Si-(CH2)2-NH2 is reacted with the surface OSiOH group of the substrate, followed by the reaction of succinic anhydride with the amine to create an amide bond and free OH, where the growth of the nucleic acid chain is supported. Cleavage includes gas cleavage with ammonia or methylamine. In some examples, cleavage includes linker cleavage with electrically generated reagents such as acids or bases. In some examples, after release from the surface, the polynucleotide is assembled into a larger polynucleotide, which is then sequenced and decoded to extract the stored information.

[0096]

[0139] The surfaces described herein can be reused after cleavage of polynucleotides to support further polynucleotide synthesis cycles. For example, linkers can be reused without additional processing / chemical modification. In some examples, linkers are non-covalently bonded to the substrate surface or polynucleotide. In some embodiments, linkers remain attached to the polynucleotide after cleavage from the surface. In some embodiments, linkers include reversible covalent bonds, such as esters, amides, ketals, β-substituted ketones, heterocyclic groups, or other groups capable of reversible cleavage. Such reversible cleavage reactions are controlled in some examples by adding or removing reagents or by electrochemical processes controlled by electrodes. Optionally, chemical linkers or chemical groups bonded to the surface are regenerated after several cycles to restore reactivity and eliminate the formation of undesirable byproducts on such linkers or chemical groups bonded to the surface.

[0097]

[0140] Alternatively, polymer synthesis can be enzymatic DNA synthesis. In some cases, enzymatic DNA synthesis uses water as the solvent, and the reagents are terminal deoxynucleotidyltransferases (TdT) or deblockers. In some cases, enzymatic DNA synthesis uses template-independent DNA polymerases, terminal deoxynucleotidyltransferases (TdT), which are proteins that have evolved to rapidly catalyze the binding of naturally occurring dNTPs. Because TdT indiscriminately adds nucleotides, uncontrolled synthesis can be prevented by various techniques, such as tethering TdT, creating mutant enzymes, and using nucleotides containing reversible terminators to prevent chain elongation. TdT activity is maximized at approximately 37°C, and the enzymatic reaction is carried out in an aqueous environment.

[0098]

[0141] extraction

[0142] In some examples, polynucleotides are deprotected, cleaved, and / or eluted from the synthesizer unit 1810 and transferred to another module in the system. In some examples, polynucleotides are transferred from the synthesizer unit 1810 on a solid support. In some examples, a robotic system 1830 or fluid tubes are used to transport polynucleotides to another module in the system. The robotic system 1830 may be controlled by a controller 1835. The robotic system generally includes a system for manipulating multiple polynucleotides. In some examples, the robotic system is used to manipulate structures containing multiple polynucleotides, such as those described herein. Operations may include, in non-limiting examples, movement, storage, acquisition, handling, transfer, or any combination thereof. The robotic system may be similar to those used in semiconductor processing to move trays of wafers and chips between processing devices. The robotic system 1830 may be used to select and transfer polynucleotides between modules of the system. For example, the robotic system 1835 may include a tag reader for verifying structures in the storage unit 1815. In some examples, the robotic system 1835 includes a tag reader (e.g., an RFID reader, a barcode reader, etc.), and the structure within the storage unit 1815 includes tags (e.g., RFID tags, barcodes, etc.). Once verified, the robotic system 1830 can transport the structure to a component of the system. In addition, the robotic system 1830 can transport the structure to a precise location within the components of the system. In some examples, the robotic system may enable polynucleotides to be added to and / or removed from modules within the data storage system. In some examples, the robotic system may enable a structure containing multiple polynucleotides to be placed and / or retrieved from a location in the storage unit 1815 with an identifiable layout. The robotic system 1830 may be controlled using a controller 1835, as further described herein.

[0099]

[0143] In some examples, one or more droplets containing polynucleotides are transferred from the synthesizer unit 1810 to the storage unit 1815. In some examples, some or all of the polynucleotides synthesized on a solid support are transferred to a structure for storage. The structure may have various shapes and sizes, such as those described herein (e.g., Figures 9A-9I). The structure may further include a tag, such as those described herein (e.g., Figures 9H-9I). The tag may include an RFID tag. In some examples, the synthesizer unit 1810 is connected to or has the same extent as a system or platform for biomolecule extraction or storage. Thus, in some examples, the polynucleotides are extracted using a system or platform including flow cells directed to maximize the recovery of the fluid containing the polynucleotides. The extracted polynucleotides may be recovered into the structure for subsequent storage. In some examples, an intermediate storage chamber is used to collect polynucleotides from a specific flow cell (or a set of flow cells) or a flow cell block (or a set of flow cell blocks). In some cases, polynucleotides are transferred from one intermediate storage chamber to another.

[0100]

[0144] The system may further include one or more mechanisms for transferring substrates to and from the system, generally as part of a larger platform for biomolecular storage. Such a platform may generally include one or more components such as flow cells, reservoirs, and pumps containing cavities for holding substrates containing biomolecules, as well as any other suitable components of the system described herein. The platform may further include a device that includes at least one logic element for performing one or more operations within the platform. In some examples, one or more operations are performed based on sensor data from one or more components of the biomolecular extraction system. In some examples, the at least one logic element comprises a programmable logic controller (PLC), a programmable logic array (PLA), a programmable array logic (PAL), a general-purpose logic array (GLA), a composite programmable logic determination (CPLD), a field-programmable gate array (FPGA), or an application-specific integrated circuit (ASIC). Such a device may communicate with or have the same extent as the controller of the system for biomolecular extraction, as described herein.

[0101]

[0145] The devices within the platform can perform one or more actions, such as (i) determining the timing of opening and closing one or more valves connecting one or more components; (ii) adjusting one or more parameters of a biomolecular extraction system, including one or more parameters such as the liquid filling rate, liquid volume, liquid, liquid discharge rate, capillary count, one or more angles of the flow cell relative to the horizontal plane, or any combination thereof; (iii) determining the recovery efficiency of the liquid discharged from the cavity containing biomolecules; or (iv) any combination thereof. In some examples, one or more metrics, parameters, sensor data, or any combination thereof may be displayed in real time on the user interface. In some examples, one or more metrics, parameters, sensor data, or any combination thereof may be analyzed in real time to detect system anomalies. In some examples, the anomaly results in a system failure. In some examples, the controller performs one or more actions in response to the system failure. In some examples, the actions include one or more of the following: logging the failure, discharging the contents of the flow cell or flow cell block into waste, resetting the flow cell or flow cell block, and marking the flow cell or flow cell block as faulty (e.g., preventing further synthesis in this device). In some examples, one or more metrics, parameters, sensor data, or any combination thereof may be stored in the cloud, random access memory, hard disk drives, solid-state drives, flash memory devices, or any variation thereof. In some examples, the device is communicatively coupled to cloud computing resources that may be used to perform any of the operations provided herein. In some examples, the systems provided herein include a graceful failover system.

[0102]

[0146] In some examples, the platform includes a mechanism for transferring the substrate from the flow cell to another processing device, such as a substrate reaction device (e.g., a synthesis module, sequencing module, or amplification module for incubating the substrate with reactants under reaction conditions), a washing device, a scanning device, or any combination thereof. Such a mechanism may also be provided to move the substrate from a printing station (e.g., inkjet printing) to a cavity in the flow cell. The transfer mechanism may include, but is not limited to, a robotic arm that can be controlled by an apparatus and / or controller as described herein. In some examples, the transfer robot is mounted on the platform of the apparatus used for synthesis. The transfer robot may include a base, an arm movably mounted on the base, and a gripping element attached to the arm, adapted to grip the substrate during transport. The element for gripping the substrate may be, for example, a movable finger-like projection. In one embodiment, during use, the robotic arm is operated so that the substrate is gripped by the gripping element. The robotic arm is moved to transfer the substrate from the printing device to the flow cell. The substrate can be positioned using other components, such as motors, pistons, conveyors, cranks, levers, etc., which will be apparent to those skilled in the art in consideration of this disclosure. As described above, in some examples the substrate can be positioned in a substrate holder or lift mechanism within the chamber of a flow cell. In some examples the holder can be adapted to be movable in order to properly position the substrate.

[0103]

[0147] Methods for biomolecule extraction may use the devices, systems, platforms, or any combination thereof provided herein. In some examples, the method for biomolecule extraction uses a flow cell described herein. In some examples, methods are developed to optimize one or more conditions or parameters of the device, system, or platform including the flow cell in order to maximize the fluid recovered from the cavity of the flow cell. In some examples, methods may be developed to achieve maximum extraction of liquid from the cavity by changing process parameters such as angles α and β as defined herein, or the discharge flow rate (e.g., air flow rate for displacing the fluid from the cavity of the flow cell). In some examples, methods may be developed to demonstrate the relationship between the volume of liquid recovered and the number of liquid capillaries (e.g., water, extraction reagents). In some examples, methods may be developed to apply them to reagents used for biomolecule extraction, such as oligomer extraction from a surface. The system may include a flow cell including a cavity. In some examples, the cavity includes a plane substantially parallel to a physical force such as gravity. In some examples, the flow cell includes a first opening and a second opening, which can be used to fill or discharge liquid from the cavity. In some examples, the flow cell includes only two openings that can be used to fill or drain liquid from the cavity. In some examples, the flow cell includes multiple openings that can be used to fill or drain liquid from the cavity. The system may further include a reservoir for holding liquid, which can be connected to the first opening using any suitable material known in the art (e.g., PFA). The system may further include a pump that can be connected to a second opening using any suitable material known in the art (e.g., PFA).

[0104]

[0148] The method may further include introducing a liquid into the cavity of the flow cell. For example, the liquid may be a liquid contained in a reservoir, which can be introduced into the cavity via a suitable conduit. In some examples, the cavity is filled at a rate (filling rate) of about 0 to about 50 μL / s. In some examples, the filling rates are about 0 to 0.5, 0 to 1, 0 to 2.5, 0 to 5, 0 to 10, 0 to 15, 0 to 20, 0 to 25, 0 to 30, 0 to 50, 0.5 to 1, 0.5 to 2.5, 0.5 to 5, 0.5 to 10, 0.5 to 15, 0.5 to 20, 0.5 to 25, 0.5 to 30, 0.5 to 50, 1 to 2.5, 1 to 5, 1 to 10, 1 to 15, 1 to 20, 1 to 25, 1 to 30, 1 to 50, 2.5 The filling rates are 5, 2.5-10, 2.5-15, 2.5-20, 2.5-25, 2.5-30, 2.5-50, 5-10, 5-15, 5-20, 5-25, 5-30, 5-50, 10-15, 10-20, 10-25, 10-30, 10-50, 15-20, 15-25, 15-30, 15-50, 20-25, 20-30, 20-50, 25-30, 25-50, or 30-50 uL / s. In some examples, the filling rates are approximately 0, 0.5, 1, 2.5, 5, 10, 15, 20, 25, 30, or 50 uL / s. In some cases, the filling rate is at least approximately 0, 0.5, 1, 2.5, 5, 10, 15, 20, 25, or 30 uL / s. In some cases, the filling rate is at most approximately 0.5, 1, 2.5, 5, 10, 15, 20, 25, 30, or 50 uL / s. In some cases, the volume of liquid filling the cavity is approximately 0 to 1000 mL.In some examples, the volume is approximately 0-100, 0-200, 0-300, 0-400, 0-500, 0-600, 0-700, 0-800, 0-900, 0-1,000, 100-200, 100-300, 100-400, 100-500, 100-600, 100-700, 100-800, 100-900, 100-1,000, 200-300, 200-400, 200-500, 200-600, 200-700, 200-800, 200-900, 200-1,000, 300-400, 300 The available sizes are ~500, 300~600, 300~700, 300~800, 300~900, 300~1000, 400~500, 400~600, 400~700, 400~800, 400~900, 400~1000, 500~600, 500~700, 500~800, 500~900, 500~1000, 600~700, 600~800, 600~900, 600~1000, 700~800, 700~900, 700~1000, 800~900, 800~1000, or 900~1000 mL. In some cases, the volume is approximately 0, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 mL. In some cases, the volume is at least approximately 0, 100, 200, 300, 400, 500, 600, 700, 800, or 900 mL. In some cases, the volume is at most approximately 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 mL.

[0105]

[0149] The method may further include draining the liquid into the cavity of the flow cell. For example, the liquid can be drained from the cavity using a pump, which can be used to introduce air into the cavity through a suitable conduit and drain the liquid from the cavity. In some examples, the cavity is evacuated at a rate (discharge rate) of about 0 to about 50 μL / s. In some examples, the discharge rates are approximately 0-0.5, 0-1, 0-2, 0-5, 0-7, 0-10, 0-15, 0-20, 0-25, 0-30, 0-50, 0.5-1, 0.5-2, 0.5-5, 0.5-7, 0.5-10, 0.5-15, 0.5-20, 0.5-25, 0.5-30, 0.5-50, 1-2, 1-5, 1-7, 1-10, 1-15, 1-20, 1-25, 1-30, 1-50, 2-5, 2-7, 2-10 , 2-15, 2-20, 2-25, 2-30, 2-50, 5-7, 5-10, 5-15, 5-20, 5-25, 5-30, 5-50, 7-10, 7-15, 7-20, 7-25, 7-30, 7-50, 10-15, 10-20, 10-25, 10-30, 10-50, 15-20, 15-25, 15-30, 15-50, 20-25, 20-30, 20-50, 25-30, 25-50, or 30-50 uL / s. In some examples, the discharge rate is approximately 0, 0.5, 1, 2, 5, 7, 10, 15, 20, 25, 30, or 50 uL / s. In some examples, the discharge rate is at least approximately 0, 0.5, 1, 2, 5, 7, 10, 15, 20, 25, or 30 uL / s. In some examples, the discharge rate is at most approximately 0.5, 1, 2, 5, 7, 10, 15, 20, 25, 30, or 50 uL / s.

[0106]

[0150] The method may further include determining the amount of liquid recovered from the cavity. In some examples, determining the amount of liquid recovered from the cavity includes comparing the properties of the liquid after it has been drained from the cavity with the properties of the liquid before the cavity was filled with the liquid. In some examples, the properties include weight, density, or volume. Thus, in some examples, determining the amount of liquid recovered from the cavity includes comparing the weight of the liquid in the reservoir after it has been drained from the cavity with the properties of the liquid before the cavity was filled with the liquid. In some examples, determining the amount of liquid recovered from the cavity may further include determining the recovery efficiency. Recovery efficiency may be determined by the percentage, ratio, or ratio of the amount of liquid drained from and recovered from the cavity to the amount of liquid that filled the cavity. In some examples, one or more sensors are configured to measure the degree of extraction. In some examples, recovery efficiency may be measured using concentration. For example, the concentration of biomolecules initially present in the cavity and / or the acquired concentration may be measured by spectrophotometric means (e.g., A260 / A280 reading), fluorescence, secondary reaction, or any other suitable method known in the art.

[0107]

[0151] A method for determining one or more conditions for recovering the maximum amount of liquid from the cavity in the flow cell may further include adjusting the parameters of the system. In some examples, the parameters include the liquid filling rate, the liquid volume, the liquid, the liquid discharge rate, the number of capillaries, one or more angles of the flow cell with respect to the horizontal plane, or any combination thereof. In some examples, one or more angles include angles with respect to a horizontal xy plane. In some examples, the horizontal xy plane is perpendicular to a physical force such as gravity. In some examples, one or more angles of the flow cell are angles α, β, or both, as defined herein. In some examples, adjusting one or more angles of the flow cell includes adjusting angle α, which can be adjusted between 0° and 90°. In some examples, adjusting one or more angles of the flow cell includes adjusting angle β, which can be adjusted between 0° and 45°. In some examples, the filling rate is adjusted between 0 and 50 μL / s. In some examples, the discharge rate is adjusted between 0 and 50 μL / s. In some examples, the volume of the liquid is adjusted between 0 mL and 1000 mL. In some examples, the capitalized number is adjusted to a value between 0 and 1 (e.g., 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, etc.). Once the parameters or conditions are adjusted, one or more operations in the method provided herein can be repeated to determine one or more conditions for recovering the maximum amount of liquid from the cavity of the flow cell.

[0108]

[0152] Furthermore, this specification provides methods for extracting biomolecules. In some examples, a flow cell, system, or platform is deployed once one or more conditions for maximizing the liquid from the cavity of the flow cell are determined. Such methods may include providing a system comprising a flow cell having a cavity, a reservoir for holding the liquid, and a pump. In some examples, the flow cell is oriented such that the plane of the cavity is substantially parallel to a bodily force such as gravity. In some examples, the flow cell is oriented such that α > 0°, β > 0°, or both. In some examples, the flow cell is oriented such that α ≤ 90°, β ≤ 45°, or both. In some examples, the flow cell is oriented such that 0° < α ≤ 90°, 0° < β ≤ 45°, or both.

[0109]

[0153] A method for extracting biomolecules may include providing a substrate. In some examples, the substrate comprises multiple biomolecules. In some examples, the extraction of biomolecules includes cleaving the biomolecules and transferring the biomolecules to a chamber. In some examples, the substrate comprises one or more chemical substances or molecules. The molecules may include, but are not limited to, polymers (e.g., peptides, proteins, nucleic acids or their mimics, e.g., peptide nucleic acids, LNA molecules, UNA molecules), polysaccharides, phospholipids, or any combination thereof, and the polymers may be heteropolymers or homopolymers. In some examples, the substrate comprises cell or tissue sections stably immobilized therein. In some examples, the substrate is manually placed in the cavity of the flow cell. In some examples, the substrate is placed in the cavity of the flow cell using an automated system. The automated system may include, but is not limited to, the transfer mechanisms described herein (e.g., robotic systems such as robotic arms).

[0110]

[0154] The method may further include exposing a substrate within the cavity of a flow cell to a fluid. In some examples, the method includes introducing a liquid into the cavity of a flow cell. In some examples, the liquid is introduced through a first opening of the flow cell or through a first number of openings of the flow cell. For example, the fluid may be filled through a single hole located at the bottom of the flow cell, thus introducing the liquid into the cavity of the flow cell and exposing the substrate within the cavity to the liquid. In some examples, the method includes removing the liquid from the cavity after introducing the liquid into the cavity of the flow cell, such as removing excess liquid through a second opening. In some examples, the liquid is removed from the cavity through a single hole located at the top of the flow cell. In some examples, the liquid is introduced into the cavity or the liquid is removed from the cavity through multiple holes. In some examples, the cavity is filled at a rate (filling rate) of about 0 to about 50 μL / s, as described herein. In some examples, the volume of liquid filling the cavity is about 0 to 1000 mL, as described herein. In some examples, the cavities may be filled with a first fluid followed by a second fluid, and thus the substrate may be exposed to the first fluid followed by the second fluid. In some examples, the fluid can be used to wash the substrate. In some examples, the fluid contains a reagent that can react with biomolecules on the substrate. In some examples, the biomolecules include nucleic acid molecules or polynucleotides. In some examples, the fluid contains a fluid for coupling, capping, oxidation, sulfidation, deblocking, or washing the substrate or biomolecules on the substrate. In some examples, the fluid is used to remove or cleave biomolecules on the surface of the substrate. The fluid may include, in non-limiting examples, water, acetone, acetonitrile, methanol, ethanol, isopropyl alcohol, or tert-butylamine. In some examples, the fluid contains tert-butylamine, water, and methanol in ratios such as about 1:1:1, 1:1:2, 1:2:1, 2:1:1, 1:2:2, 2:1:2, or 2:2:1.

[0111]

[0155] The fluid may have one or more properties suitable for use in the flow cell devices or systems described herein. These one or more properties may include, in non-limiting examples, surface tension, viscosity, density, vapor pressure, capillary action, cavitation, specific gravity, specific volume, temperature, or pressure. In some examples, the fluid has a surface tension of about 0.001 to 0.1 N / m. In some examples, the fluid is approximately 0.001-0.002, 0.001-0.005, 0.001-0.01, 0.001-0.02, 0.001-0.03, 0.001-0.04, 0.001-0.05, 0.001-0.06, 0.001-0.07, 0.001-0.08, 0.001-0.09, 0.001-0.1, 0.002-0.005, 0.002-0.01, 0.002-0.02, 0.002-0.03, 0.002-0.04, 0.002 ~0.05, 0.002~0.06, 0.002~0.07, 0.002~0.08, 0.002~0.09, 0.002~0.1, 0.005~0.01, 0.005~0.02, 0.005~0.03, 0.005~0.04, 0.005~0.05, 0.005~0.06, 0.005~0.07, 0.005~0.08, 0.005~0.09, 0.005~0.1, 0.01~0.02, 0.01~0.03, 0.01~0.04, 0.01~0.05 , 0.01~0.06, 0.01~0.07, 0.01~0.08, 0.01~0.09, 0.01~0.1, 0.02~0.03, 0.02~0.04, 0.02~0.05, 0.02~0.06, 0.02~0.07, 0.02~0.08, 0.02~0.09, 0.02~0.1, 0.03~0.04, 0.03~0.05, 0.03~0.06, 0.03~0.07, 0.03~0.08, 0.03~0.09, 0.03~0.1, 0.04~0.05, It has a surface tension of 0.04~0.06, 0.04~0.07, 0.04~0.08, 0.04~0.09, 0.04~0.1, 0.05~0.06, 0.05~0.07, 0.05~0.08, 0.05~0.09, 0.05~0.1, 0.06~0.07, 0.06~0.08, 0.06~0.09, 0.06~0.1, 0.07~0.08, 0.07~0.09, 0.07~0.1, 0.08~0.09, 0.08~0.1, or 0.09~0.1 N / m.In some examples, the fluid has a surface tension of approximately 0.001, 0.002, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, or 0.1 N / m. In some examples, the fluid has a surface tension of at least approximately 0.001, 0.002, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, or 0.09 N / m. In some examples, the fluid has a surface tension of up to approximately 0.002, 0.005, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, or 0.1 N / m. In some examples, the fluid is approximately 1 × 10⁻⁶. -3 ~5×10 -3 It has a viscosity of Pa s. In some examples, the fluid is 0.5 × 10 -3 ~1 × 10 -3 , 0.5 × 10 -3 ~1.5×10 -3 , 0.5 × 10 -3 ~2×10 -3 , 0.5 × 10 -3 ~2.5×10 -3 , 0.5 × 10 -3 ~3×10 -3 , 0.5 × 10 -3 ~3.5×10 -3 , 0.5 × 10 -3 ~4×10 -3 , 0.5 × 10 -3 ~4.5×10 -3 , 0.5 × 10 -3 ~5×10 -3 , 1 x 10 -3 ~1.5×10 -3 , 1 x 10 -3 ~2×10 -3 , 1 x 10 -3 ~2.5×10 -3 , 1 x 10 -3 ~3, 1×10 -3 ~3.5×10 -3 , 1 x 10 -3 ~4×10 -3 , 1 x 10 -3 ~4.5×10 -3 , 1 x 10 -3 ~5×10 -3 , 1.5×10-3 ~2×10 -3 、1.5×10 -3 ~2.5×10 -3 、1.5×10 -3 ~3×10 -3 、1.5×10 -3 ~3.5×10 -3 、1.5×10 -3 ~4×10 -3 、1.5×10 -3 ~4.5×10 -3 、1.5×10 -3 ~5×10 -3 、2×10 -3 ~2.5×10 -3 、2×10 -3 ~3×10 -3 、2×10 -3 ~3.5×10 -3 、2×10 -3 ~4×10 -3 、2×10 -3 ~4.5×10 -3 、2×10 -3 ~5×10 -3 、2.5×10 -3 ~3×10 -3 、2.5×10 -3 ~3.5×10 -3 、2.5×10 -3 ~4×10 -3 、2.5×10 -3 ~4.5×10 -3 、2.5×10 -3 ~5×10 -3 、3×10 -3 ~3.5×10 -3 、3×10 -3 ~4×10 -3 、3×10 -3 ~4.5×10 -3 、3×10 -3 ~5×10 -3 、3.5×10 -3 ~4×10 -3 、3.5×10 -3 ~4.5×10 -3 、3.5×10 -3 ~5×10 -3 、4×10 -3 ~4.5×10 -3 、4×10 -3~5×10 -3 , or 4.5 × 10 -3 ~5×10 -3 It has a viscosity of Pa s. In some examples, the fluid is approximately 0.5 × 10⁻⁶. -3 , 1 x 10 -3 , 1.5×10 -3 , 2×10 -3 , 2.5×10 -3 , 3 x 10 -3 , 3.5×10 -3 , 4×10 -3 , 4.5×10 -3 , or 5×10 -3 It has a viscosity of Pa s. In some examples, the fluid is approximately 0.5 × 10⁻⁶. -3 , 1 x 10 -3 , 1.5×10 -3 , 2×10 -3 , 2.5×10 -3 , 3 x 10 -3 , 3.5×10 -3 , 4×10 -3 , 4.5×10 -3 , or 5×10 -3 It has a viscosity of Pa s. In some examples, the fluid has a viscosity of at least about 0.5 × 10⁻⁶. -3 , 1 x 10 -3 , 1.5×10 -3 , 2×10 -3 , 2.5×10 -3 , 3 x 10 -3 , 3.5×10 -3 , 4×10 -3 , or 4.5 × 10 -3 It has a viscosity of Pa s. In some examples, the fluid has a viscosity of up to approximately 1 × 10⁻⁶. -3 , 1.5×10 -3 , 2×10 -3 , 2.5×10 -3 , 3 x 10 -3 , 3.5×10 -3 , 4×10 -3 , 4.5×10 -3 , or 5×10 -3 It has a viscosity of Pa s.

[0112]

[0156] The method may further include draining the liquid from the cavity of the flow cell. In some examples, the liquid in the cavity of the flow cell is drained through a first opening. In some examples, the liquid is drained from the cavity by introducing air into the cavity through, for example, a second opening. In some examples, the cavity is evacuated at a rate (drainage rate) of about 0 to 50 uL / s, as described herein. In some examples, during drainage, air is introduced through a single opening at the top of the flow cell (for example, called a second opening). Once air is introduced into the cavity, the liquid can be drained from the cavity of the flow cell. In some examples, air is introduced into the cavity or the liquid is drained from the cavity through multiple holes. In some examples, the fluid removes biomolecules from a substrate and is then drained. Thus, in some examples, the liquid contains multiple biomolecules, thereby extracting multiple biomolecules from the substrate.

[0113]

[0157] The cavity, or one or more manifolds or tubes connected to the cavity, may include a heating element or be in contact with a heating element. Therefore, in some examples, exposing a substrate placed within the cavity of the flow cell to the fluid may include heating the cavity. In some examples, discharging liquid from the cavity of the flow cell may include heating the cavity or heating one or more tubes or manifolds from which the liquid is discharged. In some examples, the heating element includes a heating jacket, a cartridge heater, or a film heater.

[0114]

[0158] The method may further include collecting a liquid containing multiple biomolecules. In some examples, the biomolecules are collected in a sample collector. In some examples, the storage stage includes the sample collector. In some examples, the sample collector includes an intermediate compartment for transferring the biomolecules for further processing or storage. In some examples, the sample collector includes a storage compartment for storing the biomolecules. The biomolecules may be stored in systems and platforms for DNA data storage, such as those provided herein (e.g., Figure 8).

[0115]

[0159] keep

[0160] The structures may have a variety of shapes and sizes, such as those described herein (e.g., Figures 9A-9I). In some examples, the structures for storing polynucleotides are located on or arranged on a plate. Structures for storing multiple polynucleotides may be of any shape or size. In some examples, the structures are substantially spherical, tubular (Figure 9A), oval, conical, cubic, rectangular, cylindrical, wedge-shaped, hexagonal prism, square pyramidal, triangular pyramidal, triangular prism, toroidal, hemispherical, helical, heart-shaped, or other shapes. In some examples, the shape is configured to allow the structure to be opened and closed to the external environment. In some examples, such closures are facilitated by welding, seals, partitions, or other mechanisms to restrict the movement of gases or other substances into or out of the structure. In some examples, the structures include holes, slots, partitions, valves, or ports for adding or removing nucleic acids, fluids, gases, or other substances to or from the structure. In some examples, the structure for storing multiple polynucleotides includes a cap and body flash-welded together (Figure 9B). In some examples, the structure for storing multiple polynucleotides includes a removable screw cap (Figure 9C). In some examples, the structure includes a septum (Figure 9D). In some examples, the structure includes two round, tablet-shaped halves that form a seal when one half is inserted into the other (9E). In some examples, the structure includes a substantially flat disc chamber with a sealable lid (Figure 9F). In some examples, the structure includes a box with an optionally fitted lid (Figure 9G). In some examples, the shape is cylindrical or disc-shaped. In some examples, a cylindrical or disc shape is preferred for automated handling and / or filing of the structure. In some examples, the chamber includes multiple vials. In some examples, the vials include borosilicate glass, plastic, metal, or other material. In some examples, the vials are placed on an alignment tray. After filling, the vial is sealed with a metal plate positioned to enclose an open vial that has been successfully post-processed and filled with polynucleotides.

[0116]

[0161] Structures for storing multiple polynucleotides may have a volume of approximately 0.1 to 1.5 mL. The volume may generally contain a certain volume of polynucleotides, a storage medium, or a combination thereof. The storage medium may be used to stabilize the polynucleotides, which may enable long-term storage. The storage medium may be, for example, a salt, an ionic liquid, glass, silica (e.g., silicone beads), or any other suitable storage solution. In some cases, the volumes are approximately 0.1mL-0.2mL, 0.1mL-0.5mL, 0.1mL-0.7mL, 0.1mL-1mL, 0.1mL-1.2mL, 0.1mL-1.5mL, 0.2mL-0.5mL, 0.2mL-0.7mL, 0.2mL-1mL, 0.2mL-1.2mL, 0.2mL-1.5mL, 0.5mL-0.7mL, 0.5mL-1mL, 0.5mL-1.2mL, 0.5mL-1.5mL, 0.7mL-1mL, 0.7mL-1.2mL, 0.7mL-1.5mL, 1mL-1.2mL, 1mL-1.5mL, or 1.2mL-1.5mL. In some cases, the volume is approximately 0.1 mL, 0.2 mL, 0.5 mL, 0.7 mL, 1 mL, 1.2 mL, or 1.5 mL. In some cases, the volume is at least approximately 0.1 mL, 0.2 mL, 0.5 mL, 0.7 mL, 1 mL, or 1.2 mL. In some cases, the volume is at most approximately 0.2 mL, 0.5 mL, 0.7 mL, 1 mL, 1.2 mL, or 1.5 mL.

[0117]

[0162] Structures for storing multiple polynucleotides may have an internal storage volume of approximately 10 to 500 microliters (μL). The internal storage volume generally contains the volume of the polynucleotides. In some examples, the internal storage volume is approximately 10 μL to 50 μL, 10 μL to 100 μL, 10 μL to 150 μL, 10 μL to 200 μL, 10 μL to 250 μL, 10 μL to 300 μL, 10 μL to 250 μL, 10 μL to 400 μL, 10 μL to 450 μL, 10 μL to 500 μL, 50 μL to 100 μL, 50 μL to 150 μL, 50 μL to 200 μL, and so on. 250μL, 50μL~300μL, 50μL~250μL, 50μL~400μL, 50μL~450μL, 50μL~500μL, 100μL~150μL, 100μL~200μL L, 100μL~250μL, 100μL~300μL, 100μL~250μL, 100μL~400μL, 100μL~450μL, 100μL~500μL, 150μL~200 μL, 150μL~250μL, 150μL~300μL, 150μL~250μL, 150μL~400μL, 150μL~450μL, 150μL~500μL, 200μL~25 0μL, 200μL~300μL, 200μL~250μL, 200μL~400μL, 200μL~450μL, 200μL~500μL, 250μL~300μL, 250μL~2 The internal storage volumes are 50 μL, 250 μL to 400 μL, 250 μL to 450 μL, 250 μL to 500 μL, 300 μL to 250 μL, 300 μL to 400 μL, 300 μL to 450 μL, 300 μL to 500 μL, 250 μL to 400 μL, 250 μL to 450 μL, 250 μL to 500 μL, 400 μL to 450 μL, 400 μL to 500 μL, or 450 μL to 500 μL. In some examples, the internal storage volume is approximately 10 μL, 50 μL, 100 μL, 150 μL, 200 μL, 250 μL, 300 μL, 250 μL, 400 μL, 450 μL, or 500 μL. In some examples, the internal storage volume is at least about 10 μL, 50 μL, 100 μL, 150 μL, 200 μL, 250 μL, 300 μL, 250 μL, 400 μL, or 450 μL.In some examples, the internal storage volume is approximately 50 μL, 100 μL, 150 μL, 200 μL, 250 μL, 300 μL, 250 μL, 400 μL, 450 μL, or 500 μL at most.

[0118]

[0163] Structures for storing multiple polynucleotides can have a diameter of approximately 1 mm to 10 mm. In some examples, the diameters are approximately 1 mm to 2 mm, 1 mm to 3 mm, 1 mm to 4 mm, 1 mm to 5 mm, 1 mm to 6 mm, 1 mm to 7 mm, 1 mm to 8 mm, 1 mm to 9 mm, 1 mm to 10 mm, 2 mm to 3 mm, 2 mm to 4 mm, 2 mm to 5 mm, 2 mm to 6 mm, 2 mm to 7 mm, 2 mm to 8 mm, 2 mm to 9 mm, 2 mm to 10 mm, 3 mm to 4 mm, 3 mm to 5 mm, 3 mm to 6 mm, 3 mm to 7 mm, 3 mm to 8 mm, and 3 mm. The diameters are approximately 1mm~9mm, 3mm~10mm, 4mm~5mm, 4mm~6mm, 4mm~7mm, 4mm~8mm, 4mm~9mm, 4mm~10mm, 5mm~6mm, 5mm~7mm, 5mm~8mm, 5mm~9mm, 5mm~10mm, 6mm~7mm, 6mm~8mm, 6mm~9mm, 6mm~10mm, 7mm~8mm, 7mm~9mm, 7mm~10mm, 8mm~9mm, 8mm~10mm, or 9mm~10mm. In some examples, the diameter is approximately 1mm, 2mm, 3mm, 4mm, 5mm, 6mm, 7mm, 7.5mm, 8mm, 9mm, or 10mm. In some examples, the diameter is at least approximately 1mm, 2mm, 3mm, 4mm, 5mm, 6mm, 7mm, 7.5mm, 8mm, or 9mm. In some examples, the diameter is up to approximately 2 mm, 3 mm, 4 mm, 5 mm, 6 mm, 7 mm, 7.5 mm, 8 mm, 9 mm, or 10 mm. As shown in Figures 15A-15B, the diameter may be a base diameter or an inner diameter. In exemplary embodiments, a structure or compartment for storing polynucleotides may include, for example, an inner diameter of approximately 2 mm and a base diameter of approximately 3 mm.

[0119]

[0164] Structures for storing multiple polynucleotides can have lengths of approximately 8mm to 30mm. In some examples, the lengths are approximately 8mm to 10mm, 8mm to 12mm, 8mm to 15mm, 8mm to 18mm, 10mm to 20mm, 8mm to 22mm, 8mm to 25mm, 8mm to 28mm, 8mm to 30mm, 10mm to 12mm, 10mm to 15mm, 10mm to 18mm, 10mm to 20mm, 10mm to 22mm, 10mm to 25mm, 10mm to 28mm, 10mm to 30mm, 12mm to 15mm, 12mm to 18mm, 12mm to 20mm, 12mm to 22mm, 12mm to 25mm, and 12mm. The lengths are approximately 10mm, 12mm, 15mm, 18mm, 20mm, 22mm, 25mm, 28mm, 30mm, 20mm, 22mm, 25mm, 28mm, 30mm, 20mm, 22mm, 20mm, 25mm, 28mm, 20mm, 30mm, 22mm, 25mm, 22mm, 28mm, 30mm, 25mm, 28mm, or 30mm. In some examples, the length is at least about 10 mm, 12 mm, 15 mm, 18 mm, 20 mm, 22 mm, 25 mm, or 28 mm. In some examples, the length is at most about 12 mm, 15 mm, 18 mm, 20 mm, 22 mm, 25 mm, 28 mm, or 30 mm. Referring to Figures 15A-15B, in exemplary embodiments, the structure or compartment for storing polynucleotides may include a length or height of about 10 mm.

[0120]

[0165] Structures for storing multiple polynucleotides can have a variety of properties. In some examples, the structure is heat-resistant, water-resistant, corrosion-resistant, airtight, biologically inert, or any combination thereof. In some examples, the structure contains metal or organic polymers. In some examples, the structure contains steel (e.g., stainless steel) or other metal alloys. In some examples, the structure contains polyethylene, polypropylene, or other polymers. In some examples, the structure contains aramid. In some examples, the structure contains meta-aramid or para-aramid. In some examples, the support is made of a flexible material. Exemplary flexible materials include, but are not limited to, modified nylon, unmodified nylon, nitrocellulose, polypropylene, and carbon fiber (such as carbon fiber composites). In some examples, the structure is made of a rigid material. Examples of rigid materials include, but are not limited to, glass, quartz glass, silicon, silicon dioxide, silicon nitride, plastics (e.g., polytetrafluoroethylene, polypropylene, polystyrene, polycarbonate, and blends thereof), and metals (e.g., steel, gold, platinum). In some examples, the structures disclosed herein may be made from materials comprising silicon, polystyrene, agarose, dextran, cellulosic polymers, polyacrylamide, polydimethylsiloxane (PDMS), glass, or any combination thereof. In some examples, the structures disclosed herein are manufactured from combinations of the materials listed herein or any other suitable materials known in the art.

[0121]

[0166] The structures described herein may include materials having a range of tensile strengths. Exemplary materials having a range of tensile strengths include, but are not limited to, nylon (70 MPa), nitrocellulose (1.5 MPa), polypropylene (40 MPa), silicon (268 MPa), polystyrene (40 MPa), agarose (1-10 MPa), polyacrylamide (1-10 MPa), and polydimethylsiloxane (PDMS) (3.9-10.8 MPa). In some examples, the material is a high-tensile-strength material (e.g., aramid). In some examples, the structures have tensile strengths of approximately 400, 600, 800, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3200, 3400, 3600, 3800, or 4000 megapascals. In some examples, the structure has a tensile strength of at least approximately 400, 600, 800, 1000, 1200, 1400, 1600, 1800, 2000, 2200, 2400, 2600, 2800, 3000, 3200, 3400, 3600, 3800, or 4000 megapascals.

[0122]

[0167] Young's modulus measures a material's resistance to elastic deformation (recoverable deformation) under load. Examples of materials with a certain range of Young's modulus stiffness include, but are not limited to, nylon (3 GPa), nitrocellulose (1.5 GPa), polypropylene (2 GPa), silicon (150 GPa), polystyrene (3 GPa), agarose (1-10 GPa), polyacrylamide (1-10 GPa), and polydimethylsiloxane (PDMS) (1-10 GPa). Structures described herein may have Young's moduli of 1-500, 1-40, 1-10, 1-5, or 3-11 GPa. The structures described herein may have a Young's modulus of approximately 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 20, 25, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 400, 500 GPa, or higher. Because the relationship between flexibility and rigidity is inverse to each other, flexible materials have a low Young's modulus and their shape changes significantly under load. In some examples, the structures described herein have a surface having flexibility of at least nylon.

[0123]

[0168] Provided herein are structures for storing polynucleotides that are corrosion-resistant. In some examples, corrosion includes uniform corrosion, pitting corrosion, crevice corrosion, galvanic corrosion, stress corrosion cracking, intergranular corrosion, temperature-induced corrosion, atmospheric corrosion, or microbial corrosion. Corrosion resistance may be conferred by the materials used to manufacture various coatings or the structure itself. Exemplary materials include metals and alloys, including iron, steel, copper, nickel, iron alloys (e.g., stainless steel, alloys with chromium such as at least 10.5% chromium, or other amounts), copper alloys, and nickel alloys. In some examples, corrosion resistance is conferred by a sacrificial anode. In some examples, the structures described herein include rare earth metals. In some examples, the structures described herein include platinum, palladium, gold, silver, rhodium, titanium, or other corrosion-resistant materials. In some examples, the coating of the structure includes glass. Glass coatings may be used in addition to, or as an alternative to, metal coatings, such as those described herein.

[0124]

[0169] The corrosion resistance of the structures described herein can be measured by various means known in the art. For example, corrosion resistance is measured using tests provided by the American Society for Testing and Materials (ASTM). Exemplary protocol categories for measuring corrosion resistance include corrosion of metals in natural air and aquatic environments, and electrochemical measurements in corrosion tests, laboratory corrosion tests, or other standard tests. Exemplary corrosion tests include, but are not limited to, salt spray, modified salt spray (acetic acid salt spray, acidified salt mist, seawater acidification, SO2 salt spray, or diluted electrolyte salt mist / dry test). The test specimen is then examined for evidence of corrosion by various means, such as weight loss, thickness loss, surface inspection (pitting / crevice), electrochemical measurements, functionality, or other methods. In exemplary measurements, the thickness loss due to corrosion of the structures described herein is expressed as MPY (mills per year, 1 mill = 0.0254 mm). In some examples, MPY = 87.6 × (metal density (g / cm³) 3 ) × sample area (cm²) 2 The formula is (time) × exposure time (hours). In some examples, the corrosion-resistant structures described herein have an MPY of less than 1 when exposed to seawater at 25°C, or an MPY of less than 0.001, 0.01, 0.05, 0.1, 0.2, 0.5, 0.7, 1, 1.2, 1.5, 2, 5, or 10. In some examples, the corrosion-resistant structures described herein have an MPY of less than 1 when exposed to a 1 percent (w / w) acetic acid solution at 100°C, or an MPY of less than 0.1, 0.5, 1, 5, 10, 20, or 50.

[0125]

[0170] Provided herein are structures for storing hermetically sealed polynucleotides. Such structures can be used to reduce or eliminate the exposure of polynucleotides to harmful / degrading elements of the external environment. In some examples, hermetically sealed refers to resistance to or impermeability to gas transfer from the environment inside the structure to the environment outside the structure, thereby reducing or eliminating contact between the gas and the stored polynucleotides, or maintaining pressure or vacuum within the structure. In some examples, hermetically sealed structures maintain a constant pressure due to welding, seals, partitions, or other sealing mechanisms. In some examples, hermetically sealed structures can maintain an internal pressure of at least 1 atm, 1.5, 2, 3, 5, 10, 20, or at least 50 atm. In some examples, hermetically sealed structures can maintain a vacuum. In some examples, an airtight structure can maintain an internal pressure (vacuum) of at least 100 torr, at least 10 torr, 1 torr, 0.1 torr, 0.01 torr, 0.001 torr, 0.0001 torr, 0.00001 torr, or at least 0.000001 torr. In some examples, the gas inside the airtight structure is an inert gas. In some examples, the inert gas is nitrogen, argon, helium, or a mixture thereof.

[0126]

[0171] Structures for storing multiple polynucleotides encoded for digital information may include components such as sensors. Sensors often provide information about the state of the structure, including environmental information. Therefore, one or more sensors can be used to monitor the data storage system. In some examples, sensors provide data on open / closed state or location, locked / unlocked state, atmospheric contents (moisture content, oxygen content), temperature, history (last open or closed), or other information. This information is optionally used to monitor the proper storage or integrity of the polynucleotides contained within the structures described herein. In some examples, sensors include one or more control polynucleotides that are sequenced to verify the fidelity of the bulk digital information encoded within the structure. In some examples, these control polynucleotides do not contain digital information. In some examples, sensors showing damage or lack of fidelity indicate errors in the digital information encoded therein. In some examples, multiple polynucleotides are reviewed by sequencing them whole or partially to determine errors. In some examples, structures containing sensors showing damage or lack of fidelity to one or more polynucleotides are flagged or discarded for review. In some examples, structures containing sensors that show damage or lack of fidelity to one or more polynucleotides are erased and rewritten with polynucleotides encoding the original or error-free digital information. In some examples, one or more sensors include temperature sensors, humidity sensors, pressure sensors, salinity sensors, light sensors, UV sensors, O2 sensors, or any combination thereof. One or more sensors may be used to monitor the environment inside, outside, or both of the structure. Sensor information may be transmitted to a controller in a data storage system to maintain the integrity of the multiple polynucleotides. In some examples, one or more sensors are integrated with tags for indexing, as described herein. In some examples, one or more sensors are integrated with RFID tags within the structure.In some cases, an RFID reader provides an alert if one or more sensors are outside a threshold.

[0127]

[0172] Structures for storing multiple polynucleotides encoded for digital information may include coatings on one or more surfaces. In some examples, the coatings reside on the inner surface of the structure (not in contact with the external environment when the structure is closed / sealed). In some examples, the coatings reside on the outer surface of the structure (exposed to the environment). In some examples, the structure includes one or more coatings, e.g., 1, 2, 3, 4, 5, or more than 5 coatings. In some examples, the coatings contain similar materials, or alternatively, at least a portion of the coatings contain different materials. Various coatings, in some examples, provide one or more properties to the surface of the structure, e.g., increased corrosion resistance, drying properties, hydrophobicity, oxygen absorption, or other properties that promote polynucleotide storage. Exemplary coatings include, but are not limited to, coatings containing plastics, synthetic polymers, glass, silica, metals, biopolymers, proteins, or other materials.

[0128]

[0173] A structure for storing polynucleotides encoded for digital information may include means for indexing its contents. The indexing means may be used to identify the structure or its contents. The indexing means may include tags, as previously stated herein. The structure may include tags, for example, as provided in Figures 9H-9I. The tags may include barcodes or RFID tags. In some examples, the tags are superficial to the structure (Figure 9H). In some examples, the tags are embedded in the structure (Figure 9I). In some examples, the tags cannot be removed. In some examples, it is impossible to remove the tags. In some examples, the tags cannot be altered. In some examples, it is impossible to alter the tags. In some examples, the tags include barcodes, RFID tags, nucleic acid barcodes, protein tags, small molecule tags, or other means for uniquely identifying the structure from other structures. In some examples, the tags are RFID tags. In some examples, the RFID tags are active RFID tags or passive RFID tags, as described herein. In some examples, the RFID tags are UHF RFID tags, HF RFID tags, or LF RFID tags, as described herein. In some examples, a database is used to store information about the contents of a structure by associating such a structure with a means for indexing (e.g., a tag). In some examples, a database is not used to store information about the contents of a structure. In such cases, a tag is used to store information about the contents of a structure. In some examples, the RFID tag is used to store metadata about the contents of a structure (e.g., multiple polynucleotides, digital information, etc.).

[0129]

[0174] The structural tag is approximately 0.1 mm. 2 ~about 5mm 2 It is possible. In some cases, the tag size is 0.1 mm. 2 ~0.5mm 2 , 0.1mm 2 ~1mm 2 , 0.1mm2 ~2mm 2 , 0.1mm 2 ~3mm 2 , 0.1mm 2 ~4mm 2 , 0.1mm 2 ~5mm 2 , 0.5mm 2 ~1mm 2 , 0.5mm 2 ~2mm 2 , 0.5mm 2 ~3mm 2 , 0.5mm 2 ~4mm 2 , 0.5mm 2 ~5mm 2 , 1mm 2 ~2mm 2 , 1mm 2 ~3mm 2 , 1mm 2 ~4mm 2 , 1mm 2 ~5mm 2 , 2mm 2 ~3mm 2 , 2mm 2 ~4mm 2 , 2mm 2 ~5mm 2 , 3mm 2 ~4mm 2 , 3mm 2 ~5mm 2 , or 4mm 2 ~5mm 2 In some cases, the tag size is 0.1 mm. 2 , 0.5mm 2 , 1mm 2 , 2mm 2 , 3mm 2 , 4mm 2 , or 5mm 2 In some cases, the tag size is at least 0.1 mm. 2 , 0.5mm 2 , 1mm 2 , 2mm 2 , 3mm 2 , or 4mm 2 In some cases, the tag size is up to 0.5 mm. 2 , 1mm 2 , 2mm2 , 3mm 2 , 4mm 2 , or 5mm 2 That is the case.

[0130]

[0175] The structural tags can have a thickness of approximately 0.1 mm to approximately 3 mm. In some examples, the thickness is 0.1 mm to 0.5 mm, 0.1 mm to 1 mm, 0.1 mm to 1.5 mm, 0.1 mm to 2 mm, 0.1 mm to 3 mm, 0.5 mm to 1 mm, 0.5 mm to 1.5 mm, 0.5 mm to 2 mm, 0.5 mm to 3 mm, 1 mm to 1.5 mm, 1 mm to 2 mm, 1 mm to 3 mm, 1.5 mm to 2 mm, 1.5 mm to 3 mm, or 2 mm to 3 mm. In some examples, the thickness is 0.1 mm, 0.5 mm, 1 mm, 1.5 mm, 2 mm, or 3 mm. In some examples, the thickness is at least 0.1 mm, 0.5 mm, 1 mm, 1.5 mm, or 2 mm. In some examples, the thickness is at most 0.5 mm, 1 mm, 1.5 mm, 2 mm, or 3 mm.

[0131]

[0176] Multiple polynucleotides in a structure can be preserved according to various preservation mechanisms. These preservation mechanisms may include, without limitation, dehydration mechanisms, ionic solvent mechanisms, salt-based containment mechanisms, glass-based containment mechanisms, or any combination thereof. In some examples, multiple polynucleotides are preserved in solution or as a solid within the structure. In some examples, the structure includes a culture medium for preserving the multiple polynucleotides. In some examples, the culture medium may be solid, liquid, gas, or any combination thereof. In some examples, the culture medium may contain an organic solvent. In some examples, the culture medium may contain water. In some examples, the culture medium may not contain water. In some examples, the culture medium may contain ethanol, isopropanol, or glycerol. In some examples, the culture medium may contain a salt solution. In some examples, the molar ratio of salt to DNA may range from about 20:1 to about 2:1. In some examples, the molar ratio depends on the molecular weight of the salt used, as well as the relative amounts of salt and DNA combined. In some examples, the molar ratio is calculated between the cations of the salt and the negatively charged phosphate groups of the DNA. In some cases, the culture medium contains a salt solution in which the molar ratio of salt cations to phosphate groups in the DNA is less than 20:1. In some cases, the salt solution is dried to produce a dried product. In some cases, the salt solution may include, as non-limiting examples, calcium chloride, calcium nitrate, calcium carbonate, calcium phosphate, magnesium chloride, magnesium sulfate, magnesium nitrate, magnesium carbonate, lanthanum chloride, lanthanum nitrate, lanthanum carbonate, lanthanum bromide, or mixtures thereof. In some cases, the salt solution may include barium(II) chloride dihydrate, calcium chloride dihydrate, copper(II) chloride anhydrous, lanthanum trichloride, magnesium chloride hexahydrate, sodium chloride, or strontium chloride hexahydrate. In some cases, the concentration of the salt solution is about 0.01 nM to about 0.1 nM.

[0132]

[0177] In some examples, the culture medium for storing multiple polynucleotides contains nanoparticles. In some examples, the nanoparticles contain silica nanoparticles. In some examples, a subset of multiple polynucleotides is encapsulated within the nanoparticles. In some examples, the nanoparticles encapsulating the polynucleotides are stored in an anhydrous or nearly anhydrous environment. In some examples, the nanoparticles contain a protective layer of silica (e.g., tetraethoxysilane). In some examples, the nanoparticles contain a co-interacting compound with the polynucleotide (e.g., N-[3-(trimethoxysilyl)propyl]-N,N,N-trimethylammonium chloride). In some examples, the nanoparticles encapsulating the polynucleotides are stored within a structure described herein. In some examples, the nanoparticle-encapsulated polynucleotides are stored on a structure such as a digital microfluidic chip. The structure may include a tag such as an RFID tag described herein. In some examples, the digital microfluidic chip enables the programmability of the fluid. In some examples, the programmability enables automated storage and / or retrieval of the polynucleotides. In some examples, each location on the digital microfluidic chip contains approximately 100GB, 500GB, 1TB, 2TB, 10TB, 20TB, 30TB, or 50TB. In some examples, each location contains approximately 50μg, 100μg, 150μg, 200μg, 250μg, 300μg, 350μg, 400μg, 450μg, 500μg, 600μg, 700μg, 800μg, 900μg, or 1000μg of nanoparticles.

[0133]

[0178] In some examples, structures for storing multiple polynucleotides are implemented as products. In some examples, the product includes a structure for storing multiple polynucleotides that encode digital information. In some examples, the structure is a highly durable structure. In some examples, the structure includes a radio frequency identification (RFID) tag, such as those described herein. In some examples, the RFID tag includes metadata about the multiple polynucleotides.

[0134]

[0179] In some examples, multiple polynucleotides are transferred to a structure in the synthesizer unit 1810. In some examples, multiple polynucleotides are transferred to the structure from a flow cell, which is part of a biomolecular extraction system in some examples. In some examples, multiple polynucleotides are transferred to a structure in the storage unit 1815. Fluid and / or electronic control of polynucleotide synthesis in the storage unit 1815 may be performed by a controller 1835. In some examples, electronics in the storage unit 1815 communicate with the controller 1835. In some examples, the polynucleotides are stored at room temperature in the storage unit 1815. In some examples, the system includes a database or file system for tracking the storage capacity in the storage unit 1815. In some examples, the database includes a control application database. In some examples, the database or file system is part of the controller 1835.

[0135]

[0180] In some examples, multiple polynucleotides are stored in a device comprising multiple compartments, a base plate, and at least one cover plate, as described herein. The polynucleotides may be transferred to the compartments using a material deposition system, for example, including a nozzle (e.g., a dispenser). The compartments may be sealed using a sealing method that utilizes the thermal expansion and / or contraction of one or more components of the system, or using, for example, a thermal adhesive. A temperature gradient between one or more components of the device, or across multiple components, may be generated by one or more temperature control systems, such as a cooling system, a heating system, or both. A temperature control system may be used to seal the device using, for example, the workflow schematically shown in Figure 20.

[0136]

[0181] Structures containing multiple polynucleotides can be stored in an identifiable layout of the storage unit 1815. The identifiable layout may include one or more racks, or variations thereof. A rack may be used to hold one or more structures containing multiple polynucleotides. In some examples, each structure is stored in a specific location in the identifiable layout. In some examples, a tag contains information about the location of the structure in the identifiable layout. For example, a tag (e.g., an RFID tag) may encode metadata that includes the location of the structure in the identifiable layout. In some examples, the rack may be located in a data center. In some examples, the rack uses a mechanical structure commonly used to mount conventional computing and data storage resources in rack units. For example, a rack may include openings adapted to support disk drives, processing blades, and / or other computer equipment. In some examples, the rack includes a tag. In some examples, the tag contains information about the structures stored in / on the rack. In some examples, the tag includes a list of the structures stored in / on the rack.

[0137]

[0182] In some examples, the storage unit 1815 may be accessed using a robotic system 1830. In some examples, the identifiable layout in the storage unit 1815 includes robot-addressable slots. Each slot may hold a structure containing multiple polynucleotides. In some examples, each slot includes a width, depth, length, or any combination thereof, to accommodate a structure containing multiple polynucleotides. In some examples, the rack includes multiple slots, each slot holding a structure containing multiple polynucleotides.

[0138]

[0183] Devices or assemblies comprising structures (e.g., compartments or containers) for storing polynucleotides as described herein are provided herein. Systems and methods for using devices or assemblies for storing polynucleotides are also provided herein. In some examples, the systems, methods, devices, or assemblies relate to long-term storage of about 20, 30, 50, or 100 years or more. The devices or assemblies may generally include one or more of a base plate, a number of compartments, and a cover plate. In some examples, methods for filling, sealing, or assembling components into devices or assemblies for use in long-term polynucleotide storage are provided herein.

[0139]

[0184] A device or assembly for storing polynucleotides may include a number of compartments. The compartments may include structures for storing polynucleotides as described elsewhere in this specification, for example, the structures shown in Figures 5–11. In some examples, the compartments have one or more tags as described herein. In some examples, the compartments include vials. In some examples, the compartments include lids. In some examples, the compartments do not include lids. Exemplary diagrams showing embodiments of structures or compartments for storing polynucleotides are provided in Figures 15A–15B. While the compartments in Figures 15A–15B are circular with a diameter, in some examples the compartments may be square, rectangular, or of other shapes.

[0140]

[0185] The compartment for storing polynucleotides may include a diameter. The diameter may be the base diameter or the inner diameter. In some examples, the inner diameter is smaller than the base diameter. The base diameter may be about 1 mm to about 10 mm, as described elsewhere in this specification. In some examples, the base diameter is about 1 mm to about 5 mm. In some examples, the inner diameter may be about 1 mm to about 10 mm, as described elsewhere in this specification. In some examples, the inner diameter is about 1 mm to about 5 mm. Referring to Figures 15A-15B, the base diameter may be about 3.2 mm and the inner diameter may be about 2.2 mm.

[0141]

[0186] The compartment for storing polynucleotides may include a height. The height (or length) may be approximately 8 mm to 15 mm, as described elsewhere in this specification. In some examples, the height is approximately 8 mm to 12 mm. In some examples, the height is less than approximately 10 mm. Referring to Figures 15A-15B, the height may be approximately 9.95 mm.

[0142]

[0187] In some cases, the partition or structure has a thickness. In some cases, the thickness is approximately 0.1 mm to approximately 2 mm. In some cases, the thickness is approximately 0.5 mm. In some cases, the thickness is 0.1 to 0.2 mm, 0.1 to 0.5 mm, 0.1 to 0.8 mm, 0.1 to 1 mm, 0.1 to 1.2 mm, 0.1 to 1.5 mm, 0.1 to 1.8 mm, 0.1 to 2 mm, 0.2 to 0.5 mm, 0.2 to 0.8 mm, 0.2 to 1 mm, 0.2 to 1.2 mm, 0.2 to 1.5 mm, 0.2 to 1.8 mm, 0.2 to 2 mm, 0.5 to 0.8 mm, 0.5 to 1 mm. The thicknesses are 0.5-1.2mm, 0.5-1.5mm, 0.5-1.8mm, 0.5-2mm, 0.8-1mm, 0.8-1.2mm, 0.8-1.5mm, 0.8-1.8mm, 0.8-2mm, 1-1.2mm, 1-1.5mm, 1-1.8mm, 1-2mm, 1.2-1.5mm, 1.2-1.8mm, 1.2-2mm, 1.5-1.8mm, 1.5-2mm, or 1.8-2mm. In some cases, the thickness is 0.1, 0.2, 0.5, 0.8, 1, 1.2, 1.5, 1.8, or 2mm. In some cases, the thickness is at least 0.1, 0.2, 0.5, 0.8, 1, 1.2, 1.5, or 1.8mm. In some examples, the thickness is up to 0.2, 0.5, 0.8, 1, 1.2, 1.5, 1.8, or 2 mm.

[0143]

[0188] Multiple polynucleotides can be stored within a compartment. Multiple polynucleotides can exist in a liquid or a gas. In some examples, multiple polynucleotides are in a liquid when they are transferred to or from a compartment. In some examples, multiple polynucleotides are in a liquid when they are stored in a compartment. In some examples, multiple polynucleotides are in a solid when they are transferred to or from a compartment. In some examples, multiple polynucleotides are in a solid when they are stored in a compartment. In an exemplary manner, multiple polynucleotides are in a solution when they are transferred to a compartment.

[0144]

[0189] In some examples, each compartment of a plurality of compartments contains a certain volume of solution containing a plurality of polynucleotides. In some examples, the volume, also called the internal storage volume, is about 10 μL to about 500 μL, as described elsewhere herein. In some examples, the volume is about 10 μL to about 200 μL. In some examples, the volume is less than about 100 μL. In some examples, the volume is about 10 μL to about 50 μL. The plurality of polynucleotides may be dried when they are stored (e.g., for long-term storage). In some examples, the solution is dried under vacuum. In some examples, the plurality of polynucleotides are in solution when they are obtained or accessed. In some examples, the plurality of polynucleotides are dissolved in solution when they are obtained.

[0145]

[0190] A compartment for storing polynucleotides may comprise a first material. The first material may be inert. More specifically, the first material may be biologically inert so as not to interact with the material in the compartment (e.g., polynucleotides). The compartment may comprise a variety of properties described herein, including but not limited to heat resistance, water resistance, corrosion resistance, or airtightness. In some examples, the material for the compartment may comprise metals or organic polymers, e.g., but not limited to those described herein. In some examples, the material has a tensile strength or Young's modulus within the range described herein. In some examples, the material comprises glass. In some examples, the material comprises silica (or silicon dioxide). In some examples, the material comprises at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% silica. In some examples, the material further comprises metal oxides, e.g., but not limited to boron oxide, sodium oxide, potassium oxide, aluminum oxide, or any combination thereof. Various types of glass that can be selected as the first material are provided, for example, in Figure 19. Referring to Figure 15A, the compartment may, in exemplary embodiments, include a borosilicate. In some examples, the compartment includes a glass vial.

[0146]

[0191] In some examples, multiple compartments are positioned on a base plate. In some examples, multiple compartments are arranged on a base plate to facilitate storage of multiple compartments or retrieval of polynucleotides from multiple compartments. For example, multiple compartments may be arranged to facilitate filling the compartments with DNA suspended in solution using a dispenser such as a nozzle or pipette, or by drying the DNA. Figure 15A provides an example showing a nozzle that may be used to deliver or remove material from a compartment.

[0147]

[0192] In some examples, dispensers (e.g., nozzles, pipettes, etc.) are part of a material loading system. A material loading system may have multiple dispensers (e.g., multiple nozzles). In some examples, dispensers for filling compartments are different from dispensers for removing the contents of compartments. In some examples, one or more dispensers for filling compartments, one or more nozzles for removing the contents of compartments, or both are part of a material loading system.

[0148]

[0193] Dispensers (e.g., nozzles, pipettes, etc.) may have a variety of dimensions suitable for use with the compartments described herein. In some examples, the dispenser width is about 0.5 to about 5 mm. In some examples, the dispenser width is 0.5 to 1, 0.5 to 2, 0.5 to 3, 0.5 to 4, 0.5 to 5, 1 to 2, 1 to 3, 1 to 4, 1 to 5, 2 to 3, 2 to 4, 2 to 5, 3 to 4, 3 to 5, or 4 to 5 mm. In some examples, the dispenser width is 0.5, 1, 2, 3, 4, or 5 mm. In some examples, the dispenser width is at least 0.5, 1, 2, 3, or 4 mm. In some examples, the dispenser width is at most 1, 2, 3, 4, or 5 mm. Referring to Figure 15A, the nozzle width is about 1.38 mm in an exemplary embodiment. However, this width may vary at least in part based on the inner diameter of the compartment. In some examples, the dispenser tip includes a diameter. The diameter may be the inner diameter (referred to as the tip orifice in Figure 15A) or the outer diameter (OD). In some examples, the diameter is approximately 0.1 mm to 1 mm. In some examples, the diameters are 0.1-0.2mm, 0.1-0.3mm, 0.1-0.4mm, 0.1-0.5mm, 0.1-0.6mm, 0.1-0.7mm, 0.1-0.8mm, 0.1-0.9mm, 0.1-1mm, 0.2-0.3mm, 0.2-0.4mm, 0.2-0.5mm, 0.2-0.6mm, 0.2-0.7mm, 0.2-0.8mm, 0.2-0.9mm, 0.2-1mm, 0.3-0.4mm, 0.3-0.5mm, 0.3-0.6mm, 0.3-0.7mm, 0.3-0. The available sizes are 8mm, 0.3-0.9mm, 0.3-1mm, 0.4-0.5mm, 0.4-0.6mm, 0.4-0.7mm, 0.4-0.8mm, 0.4-0.9mm, 0.4-1mm, 0.5-0.6mm, 0.5-0.7mm, 0.5-0.8mm, 0.5-0.9mm, 0.5-1mm, 0.6-0.7mm, 0.6-0.8mm, 0.6-0.9mm, 0.6-1mm, 0.7-0.8mm, 0.7-0.9mm, 0.7-1mm, 0.8-0.9mm, 0.8-1mm, or 0.9-1mm. In some examples, the diameter is 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1 mm.In some examples, the diameter is at least 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, or 0.9 mm. In some examples, the diameter is at most 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or 1 mm. Referring to Figure 15A, in an exemplary embodiment, the inner diameter is approximately 0.35 mm, or the outer diameter is approximately 0.7 mm. However, the inner diameter, outer diameter, or both may vary at least in part based on the dimensions of the compartment or the material being filled or removed from the compartment (e.g., the viscosity of the solution).

[0149]

[0194] In some examples, multiple compartments are arranged in an array on a base plate. An exemplary schematic diagram of the base plate is shown in Figure 16. The base plate may include multiple recessed features. In some examples, the recessed features may also be called tapped holes. The multiple recessed features of the base plate may have a diameter of about 1 mm to about 10 mm, as described elsewhere in this specification. In some examples, the multiple recessed features of the base plate have a diameter of about 1 mm to about 5 mm. In some examples, each of the multiple recessed features of the base plate has the same diameter as the base diameter of one of the compartments.

[0150]

[0195] Multiple compartments can be positioned within recessed features of the base plate. These recessed features of the base plate can form wells, including walls. In some examples, each compartment is located entirely or partially within the recessed features of the base plate. In some examples, the compartments are integrated with the base plate. In some examples, the compartments and the base plate are connected, interact, or act toward each other by mechanical or electronic means. In some examples, the compartments are separable from the base plate.

[0151]

[0196] In some cases, the walls formed by the concave features are approximately 1 mm to 10 mm in length. In some cases, the walls are 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9, 1-10, 2-3, 2-4, 2-5, 2-6, 2-7, 2-8, 2-9, 2-10, 3-4, 3-5, 3-6, 3-7, 3-8, 3-9, 3-10, 4-5, 4-6, 4-7, 4-8, 4-9, 4-10, 5-6, 5-7, 5-8, 5-9, 5-10, 6-7, 6-8, 6-9, 6-10, 7-8, 7-9, 7-10, 8-9, 8-10, or 9-10 mm in length. In some examples, the wall is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 mm long. In some examples, the wall is at least 1, 2, 3, 4, 5, 6, 7, 8, or 9 mm long. In some examples, the wall is at most 2, 3, 4, 5, 6, 7, 8, 9, or 10 mm long. For example, as shown in Figure 18, the wall may be approximately 2 mm long.

[0152]

[0197] In some examples, as shown in Figure 18, for instance, each wall of the concave feature is at least partially inclined. In some examples, each wall of the concave feature of the base plate is at least partially angled at approximately 0.1° to 1°. In some examples, each wall of the concave feature of the base plate is at least partially angled at less than 1°. In some examples, each wall of the concave feature of the base plate is partially inclined at approximately 0.1°, 0.2°, 0.3°, 0.4°, 0.5°, 0.6°, 0.7°, 0.8°, 0.9°, or 1°. In some examples, each wall of the concave feature of the base plate is at least partially inclined at approximately 0.1°, 0.2°, 0.3°, 0.4°, 0.5°, 0.6°, 0.7°, 0.8°, 0.9°, or 1°. In some examples, each wall of the concave feature of the base plate is partially inclined by up to approximately 0.1°, 0.2°, 0.3°, 0.4°, 0.5°, 0.6°, 0.7°, 0.8°, 0.9°, or 1°. In some examples, the walls of the concave feature of the base plate are 0.1°~0.2°, 0.1°~0.3°, 0.1°~0.4°, 0.1°~0.5°, 0.1°~0.6°, 0.1°~0.7°, 0.1°~0.8°, 0.1°~0.9°, 0.1°~1°, 0.2°~0.3°, 0.2°~0.4°, 0.2°~0.5°, 0.2°~0.6°, 0.2°~0.7°, 0.2°~0.8°, 0.2°~0.9°, 0.2°~1°, 0.3°~0.4°, 0.3°~0.5°, 0.3°~0.6°, 0.3°~0.7°, 0.3 The slopes are partially inclined at 0.8°, 0.3°, 0.9°, 0.3°, 1°, 0.4°, 0.5°, 0.4°, 0.6°, 0.4°, 0.7°, 0.4°, 0.8°, 0.4°, 0.9°, 1°, 0.5°, 0.6°, 0.5°, 0.7°, 0.5°, 0.8°, 0.5°, 0.9°, 1°, 0.6°, 0.7°, 0.6°, 0.8°, 0.6°, 0.9°, 1°, 0.7°, 0.8°, 0.8°, 1°, or 0.9° to 1°.

[0153]

[0198] In some examples, each wall between the concave features has a thickness of approximately 0.5 mm to approximately 3 mm. In some examples, the thickness is 0.5–1, 0.5–1.5, 0.5–2, 0.5–2.5, 0.5–3, 1–1.5, 1–2, 1–2.5, 1–3, 1.5–2, 1.5–2.5, 1.5–3, 2–2.5, 2–3, or 2.5–3 mm. In some examples, the thickness is approximately 0.5, 1, 1.5, 2, 2.5, or 3 mm. In some examples, the thickness is at least approximately 0.5, 1, 1.5, 2, or 2.5 mm. In some examples, the thickness is at most approximately 1, 1.5, 2, 2.5, or 3 mm. Referring to Figure 18, in an exemplary embodiment, the thickness is approximately 1.3 mm.

[0154]

[0199] The concave features may have a pitch distance. In some examples, the concave features of the base plate have a uniform pitch. In some examples, the pitch distance is approximately 1 to 15 mm. In some examples, the pitch distance is 1-2 mm, 1-3 mm, 1-4 mm, 1-5 mm, 1-6 mm, 1-7 mm, 1-8 mm, 1-9 mm, 1-10 mm, 1-12 mm, 1-15 mm, 2-3 mm, 2-4 mm, 2-5 mm, 2-6 mm, 2-7 mm, 2-8 mm, 2-9 mm, 2-10 mm, 2-12 mm, 2-15 mm, 3-4 mm, 3-5 mm, 3-6 mm, 3-7 mm, 3-8 mm, 3-9 mm, 3-10 mm, 3-12 mm, 3-15 mm, 4-5 mm, 4-6 mm, 4-7 mm, 4-8 The pitch distances are 1, 2, 3, 4, 10, 4, 12, 4, 15 mm, 5, 6, 7, 8, 9, 10, 12, 5, 15 mm, 6, 7, 8, 6, 9, 6, 10, 6, 12, 6, 15 mm, 7, 8, 7, 9, 7, 10, 7, 12, 7, 15 mm, 8, 9, 8, 10, 8, 12, 8, 15 mm, 9, 10, 9, 12, 9, 15 mm, 10, 12, 10, 15 mm, or 12-15 mm. In some examples, the pitch distance is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, or 15 mm. In some examples, the pitch distance is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 12 mm. In some examples, the pitch distance is at most 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, or 15 mm. Referring to Figure 16 or Figure 18, in an exemplary embodiment, the pitch distance is approximately 4.5 mm.

[0155]

[0200] The base plate may include a second material. In some examples, the second material is biologically inert. In some examples, the second material includes a metal. In some examples, the second material includes a high specific heat capacity material. High specific heat capacity materials may generally have a specific heat capacity of about 0.5 J / k-°C to 2.5 J / k-°C. In some examples, the specific heat capacity is about 0.5, 1, 1.5, or 2 J / k-°C. In some examples, the specific heat capacity is at least about 0.5, 1, 1.5, or 2 J / k-°C. In some examples, the specific heat capacity is at most about 0.5, 1, 1.5, or 2 J / k-°C. In some examples, the specific heat capacity is about 0.5 to 1, 0.5 to 1.5, or 0.5 to 2, 1 to 1.5, 1 to 2, or 1.5 to 2 J / k-°C. High specific heat capacity materials may include, as non-limiting examples, aluminum, beryllium, or magnesium. In some examples, the base plate comprises a metal mounting plate, a cooling plate, or both.

[0156]

[0201] The device may further include a cover plate. In some examples, the cover plate includes a plurality of recessed features. The plurality of recessed features (also called tapped holes) may correspond to the same positions as the plurality of recessed features on the base plate. In some examples, the plurality of recessed features on the base plate are arranged in an array, and the plurality of recessed features on the cover plate are arranged in an array. Each of the plurality of recessed features on the cover plate may fit into a plurality of compartments, for example, as provided in Figure 18. In some examples, the diameter of each recessed feature on the cover plate is the same as or similar to the base diameter of each compartment. In some examples, the diameter of each recessed feature on the cover plate is the same as or similar to the base diameter of each compartment at approximately 15°C to 30°C or approximately room temperature.

[0157]

[0202] The exemplary schematic diagram in Figure 16 may also correspond to a cover plate. The multiple recessed features of the cover plate may have a diameter of about 1 mm to about 10 mm, as described elsewhere in this specification. In some examples, the multiple recessed features of the cover plate have a diameter of about 1 mm to about 5 mm. In some examples, each of the multiple recessed features of the cover plate has a diameter equal to or similar to the base diameter of one of the multiple sections. The recessed features of the cover plate may have a pitch distance, as similarly described for the base plate. In some examples, the recessed features of the cover plate have a uniform pitch. In some examples, the pitch distance is about 1 to about 15 mm, as described. Referring to Figure 16 or Figure 18, in the exemplary embodiment, the diameter of each recessed feature of the cover plate is about 3.195 mm, or the pitch distance is about 4.5 mm, or both.

[0158]

[0203] In some examples, the multiple recessed features of the cover plate form wells including walls. In some examples, each wall of the recessed feature is at least partially inclined, as shown in Figure 18, for example. In some examples, each wall of the recessed feature of the base plate is at least partially angled at about 0.1° to 1°, as described herein. In some examples, each wall of the recessed feature of the base plate is at least partially angled at less than 1°, as described herein. In some examples, the walls of the recessed feature of the cover plate are partially angled at the same or similar angle as the walls of the recessed feature of the base plate. In some examples, each wall between the recessed features of the cover plate has a thickness of about 0.5 mm to about 3 mm, as described herein. In some examples, the thickness of the walls between the recessed features of the cover plate is the same as or similar to the thickness of the walls between the recessed features of the base plate. Referring to Figure 18, in exemplary embodiments, the walls may be partially inclined at less than 0.5°, and the thickness of the walls between the recessed features of the cover plate is about 1.3 mm, or both. Furthermore, the wall formed by the concave feature of the cover plate may have a length of about 1 mm to about 10 mm, as described herein. In some examples, the wall formed by the concave feature of the cover plate may be higher than the wall formed by the concave feature of the base plate. The thickness of the upper surface of the concave feature of the cover plate may be about 0.005 to about 0.1 mm. In some examples, the thickness of the top surface may be approximately 0.005–0.01, 0.005–0.02, 0.005–0.05, 0.005–0.08, 0.005–0.1, 0.01–0.02, 0.01–0.05, 0.01–0.08, 0.01–0.1, 0.02–0.05, 0.02–0.08, 0.02–0.1, 0.05–0.08, 0.05–0.1, or 0.08–0.1 mm. In some examples, the thickness of the top surface is 0.005, 0.01, 0.02, 0.05, 0.08, or 0.1 mm. In some examples, the thickness of the top surface is at least 0.005, 0.01, 0.02, 0.05, or 0.08 mm. In some examples, the thickness of the top surface is a maximum of 0.01, 0.02, 0.05, 0.08, or 0.1 mm.Referring to Figure 18, in an exemplary embodiment, the wall formed by the concave feature of the cover plate is approximately 5.5 mm thick, and the thickness of the cover plate on the upper surface of the concave feature is approximately 0.021 mm, or both.

[0159]

[0204] The cover plate may be arranged on multiple compartments such that multiple compartments can be fully or partially accommodated by the walls of the cover plate, the walls of the base plate, or both. In some examples, the walls of each recessed feature of the cover plate and the outer walls of the compartments are separated by approximately 1 μm to approximately 10 μm. In some examples, the walls of each recessed feature of the cover plate and the outer walls of the compartments are separated by approximately 1 μm, 2 μm, 3 μm, 4 μm, 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, or 10 μm. In some examples, the walls of each recessed feature of the cover plate and the outer walls of the compartments are separated by at least approximately 1 μm, 2 μm, 3 μm, 4 μm, 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, or 10 μm. In some examples, the walls of each recessed feature and the outer walls of each compartment of the cover plate are separated by a maximum of approximately 1 μm, 2 μm, 3 μm, 4 μm, 5 μm, 6 μm, 7 μm, 8 μm, 9 μm, or 10 μm. In some examples, the walls of each recessed feature and the outer walls of each compartment of the cover plate are separated by approximately 1-2, 1-3, 1-4, 1-5, 1-6, 1-8, 1-10, 2-3, 2-4, 2-5, 2-6, 2-8, 2-10, 3-4, 3-5, 3-6, 3-8, 3-10, 4-5, 4-6, 4-8, 4-10, 5-8, 5-10, 6-8, 6-10, 7-9, 7-10, 8-10, or 9-10 μm. In some cases, the walls of the concave features of the cover plate and the outer walls of the compartments are separated by a distance of 5 μm or less at approximately 15°C to 30°C or at approximately room temperature.

[0160]

[0205] The cover plate may contain a third material. In some examples, the third material is biologically inert. In some examples, the third material contains metal. In some examples, the third material contains low emissivity. In some examples, the third material contains stainless steel. In some examples, the third material has a coefficient of thermal expansion (CTE) of approximately 10 μm / m-℃ to 25 μm / m-℃. In some examples, the third material has a CTE of approximately 15 μm / m-℃ to 20 μm / m-℃. In some examples, the third material has a CTE of 17 μm / m-℃. In some examples, the third material has a CTE of approximately 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 μm / m-℃. In some examples, the third material has a CTE of approximately 10-12, 10-15, 10-18, 10-20, 10-25, 12-15, 12-18, 12-20, 12-25, 15-18, 15-20, 15-22, 15-25, 18-20, 18-22, 18-25, 20-22, or 20-25 μm / m-℃. In some examples, the CTE of the first material in the compartment, the second material in the base plate, or both is smaller than the CTE of the third material in the cover plate. In some examples, the second material in the base plate and the third material in the cover plate have the same or similar CTEs. In some examples, the first material in multiple compartments has a lower CTE than the third material in the cover plate.

[0161]

[0206] In some examples, the third material has a thermal conductivity of approximately 10 W / mK to approximately 25 W / mK. In some examples, the third material has a thermal conductivity of approximately 15 W / mK to approximately 20 W / mK. In some examples, the third material has a thermal conductivity of 16.2 W / mK. In some examples, the third material has a thermal conductivity of approximately 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 W / mK. In some examples, the third material has a thermal conductivity of approximately 10-12, 10-15, 10-18, 10-20, 10-25, 12-15, 12-18, 12-20, 12-25, 15-18, 15-20, 15-22, 15-25, 18-20, 18-22, 18-25, 20-22, or 20-25 W / mK.

[0162]

[0207] In some examples, the third material has a specific heat capacity of approximately 0.1 to 1 J / g-°C. In some examples, the third material has a specific heat capacity of approximately 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, or approximately 1 J / g-°C. In some examples, the third material contains a specific heat capacity of approximately 0.1-0.2, 0.1-0.5, 0.1-0.8, 0.1-1, 0.2-0.5, 0.2-0.8, 0.2-1, 0.3-0.5, 0.3-0.8, 0.3-1, 0.4-0.6, 0.4-0.8, 0.4-1, 0.5-0.8, 0.5-1, 0.6-0.8, 0.6-1, 0.7-0.9, 0.7-1, 0.8-1, or 0.9-1 J / g-℃. In some examples, the third material contains a specific heat capacity of 0.5 J / g-℃.

[0163]

[0208] A device or assembly may include at least one cover plate. In some examples, a device includes two or more cover plates. In some examples, a device includes multiple cover plates. In some examples, a device has about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 150, or 200 cover plates. In some examples, a device has at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 150, or 200 cover plates. In some examples, the device may have up to approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 150, or 200 cover plates. In some examples, the device may include approximately 1-10, 1-50, 1-100, 10-50, 10-80, 10-100, 40-80, 40-100, 40-120, 50-80, 50-100, 50-120, 50-200, 60-100, 60-150, 80-100, 80-120, 80-150, 80-200, 100-120, 100-150, 100-200, or 150-200 cover plates. In some examples, the device includes approximately 64 cover plates. In some examples, the device includes approximately 96 cover plates. In some examples, each compartment of a group of compartments has its own cover plate. In some examples, each cover plate covers approximately 1, 2, 3, 4, 5, 6, 10, 12, 16, 24, 30, 36, 40, 48, or 96 compartments of the group of compartments. In some examples, each cover plate covers at least approximately 1, 2, 3, 4, 5, 6, 10, 12, 16, 24, 30, 36, 40, 48, or 96 compartments of the group of compartments. In some examples, each cover plate covers up to approximately 1, 2, 3, 4, 5, 6, 10, 12, 16, 24, 30, 36, 40, 48, or 96 compartments of the group of compartments.In some examples, each cover plate covers approximately 1-5, 1-10, 1-12, 1-20, 1-50, 1-100, 2-5, 2-10, 2-15, 2-20, 2-50, 4-10, 4-20, 4-50, 5-10, 5-20, 5-30, 5-50, 10-15, 10-20, 10-50, 10-100, 20-40, 20-50, 20-100, 40-50, 40-100, 50-80, or 50-100 of the multiple compartments. In some examples, the cover plate covers all of the compartments of the device. In some examples, the cover plate covers all of the compartments placed on the base plate. In some examples, the device includes at least approximately two cover plates. In some examples, each of the at least two cover plates seals approximately six of the multiple compartments. In some cases, the cover plate and base plate are the same size. In some cases, the cover plate is approximately 1%, 2%, 3%, 4%, 5%, 8%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the base plate size. In some cases, the cover plate is approximately 1-5%, 2-8%, 5-10%, 8-20%, 15-30%, 20-40%, 30-50%, 40-60%, 50-70%, 60-80%, 70-90%, or 80-100% of the base plate size.

[0164]

[0209] Any dimensions of components (e.g., compartments or structures, base plates or cover plates) in a device for storing polynucleotides may include tolerances. Dimensions may be any dimensions described or illustrated herein, for example, in non-limiting examples, diameter (e.g., base diameter, inner diameter), height, length, thickness, pitch distance, or any combination thereof. In some examples, any one of the compartment dimensions may include a tolerance of approximately 1 sigma to 6 sigma. In some examples, any one of the compartment dimensions may include a tolerance of up to approximately 3 sigma. In some examples, the tolerance for compartment dimensions is approximately 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 2 to 3, 2 to 4, 2 to 5, 2 to 6, 3 to 4, 3 to 5, 3 to 6, 4 to 5, 4 to 6, or approximately 5 to 6 sigma. In some examples, the tolerance for compartment dimensions is approximately 1, 2, 3, 4, 5, or 6 sigma. In some examples, the tolerance for the dimensions of a section is at most about 1, 2, 3, 4, 5, or 6 sigma. In some examples, the dimensions include, for example, the diameter, pitch distance, length, or thickness as specified herein. In some examples, the dimensions include tolerances of about 10 μm or less. In some examples, the dimensions include tolerances of about 10, 8, 5, 4, 3, 2, or 1 μm or less. In some examples, the dimensions include tolerances of about 1-2, 1-3, 1-4, 1-5, 1-6, 1-8, 1-10, 2-3, 2-4, 2-5, 2-8, 2-10, 3-5, 3-8, 3-10, 4-8, 4-10, 5-8, 5-10, 6-10, or 8-10 μm.

[0165]

[0210] A device or assembly described herein, comprising a base plate, a plurality of compartments, and a cover plate, may be used to store information. The information may be encoded in DNA. A method for storing information may include providing a device comprising a plurality of compartments, a base plate, and a cover plate, one or more of the latter. In some examples, the plurality of compartments contain a plurality of polynucleotides. In some examples, the plurality of compartments may be a plurality of glass vials. In some examples, the base plate, the plurality of compartments, or both are not in contact with the cover plate. The plurality of compartments may be arranged in an array on the base plate (or mounting plate) to facilitate filling or drying of the material in the compartments. In some examples, the method includes transferring the plurality of polynucleotides into the compartments. In some examples, the transfer includes transferring the plurality of polynucleotides in solution. In some examples, synthetic DNA suspended in a liquid solution is transferred to the compartments using a dispenser such as a nozzle or pipette. The distribution may be part of a material deposition system, as further described herein. Thus, in some examples, the transfer includes depositing one or more droplets containing the plurality of polynucleotides using a nozzle of the deposition system.

[0166]

[0211] Each of the multiple compartments may be filled with a certain amount of solution containing multiple polynucleotides. The volume may be the same as, similar to, or less than, the internal volume of one of the compartments. Thus, in some examples, the compartments are filled with a volume of approximately 10 μL to approximately 500 μL. In some examples, the compartments are filled with a volume of approximately 10 μL to approximately 200 μL. In some examples, the volume is less than approximately 100 μL. In some examples, the volume is approximately 10 μL to approximately 50 μL.

[0167]

[0212] The contents of the compartment (e.g., DNA) can be stored in solution or as a solid. In some cases, the contents of the compartment are dried, for example, by vacuum. In some cases, by drying the contents, the dried DNA remains at the bottom of the compartment.

[0168]

[0213] The method may include positioning the cover plate above the base plate and / or multiple compartments. The cover plate may be positioned above the base plate and / or multiple compartments using a piezo stage, a vision system, a linear motor, a rotary motor, any combination thereof, or any other method known in the art.

[0169]

[0214] Methods for sealing compartments within a device or assembly provided herein may generally involve the use of temperature. Thermal seals may utilize, for example, the thermal contraction and / or thermal expansion of materials of the device components (e.g., base plates, cover plates, or compartments), or a thermal adhesive for sealing one or more components of the device.

[0170]

[0215] The method generally involves generating a temperature gradient across the components of the device to seal the contents of a compartment. In some examples, a temperature gradient is generated between a base plate and a cover plate. In some examples, the temperature gradient causes the base plate or cover plate to expand or contract. In some examples, the expansion or contraction of one or more components of the device can seal the compartment and its contents. In some examples, generating a temperature gradient involves cooling or heating the base plate and / or the compartment within it, the cover plate, or both. In some examples, generating a temperature gradient involves sequentially changing the temperature of the base plate and / or the compartment within the base plate and the cover plate. In some examples, generating a temperature gradient involves simultaneously changing the temperature of the base plate and / or the compartment within the base plate and the cover plate.

[0171]

[0216] In some examples, generating a temperature gradient involves one or more operations. An exemplary workflow of one or more operations is generally shown in Figure 20. One or more operations may be performed simultaneously or sequentially. In some examples, one or more operations include cooling the base plate. The base plate, which may include mounting plates and / or cooling plates, may be cooled in an inert atmosphere. In some examples, multiple compartments that may be placed on the base plate are also cooled. The base plate may be cooled to a predetermined temperature. In some examples, the base plate is cooled from approximately -100 degrees Celsius to approximately 100 degrees Celsius. In some examples, the base plate is cooled from approximately -50 degrees Celsius to approximately 50 degrees Celsius. In some examples, the base plate is cooled to approximately -100°C, -90°C, -80°C, -70°C, -60°C, -50°C, -40°C, -30°C, -20°C, -10°C, -5°C, 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C, or approximately 100°C. In some examples, the base plate is cooled to at least approximately -100°C, -90°C, -80°C, -70°C, -60°C, -50°C, -40°C, -30°C, -20°C, -10°C, -5°C, 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C, or approximately 100°C. In some examples, the base plate is cooled to a maximum of approximately -100°C, -90°C, -80°C, -70°C, -60°C, -50°C, -40°C, -30°C, -20°C, -10°C, -5°C, 0°C, 5°C, 10°C, 15°C, 20°C, 25°C, 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C, or approximately 100°C.In some examples, the base plate can withstand temperatures of up to approximately -100°C to -80°C, -100°C to -50°C, -100°C to -20°C, -100°C to 0°C, -80°C to -50°C, -80°C to -20°C, -80°C to 0°C, -80°C to 20°C, -50°C to -20°C, -50°C to -10°C, -50°C to 0°C, -50°C to 10°C, -50°C to 20°C, -30°C to -20°C, -30°C to - 10℃, -30℃~0℃, -30℃~10℃, -30℃~20℃, -20℃~-10℃, -20℃~-5℃, -20℃~0℃, -20℃~5℃, -20℃~10℃, -20℃~20℃, -10℃~-5℃, -10℃~0℃, -10℃~5℃, -10℃~10℃, -10℃~15℃, -10℃~20℃, -10℃~25℃, -5℃~0℃, -5℃~5℃, -5℃~10 °C, -5°C to 15°C, -5°C to 20°C, -5°C to 25°C, 0°C to 5°C, 0°C to 10°C, 0°C to 15°C, 0°C to 20°C, 0°C to 25°C, 0°C to 30°C, 5°C to 10°C, 5°C to 15°C, 5°C to 20°C, 5°C to 25°C, 5°C to 30°C, 5°C to 40°C, 10°C to 15°C, 10°C to 20°C, 10°C to 25°C, 10°C to 30°C, 10°C to 40°C, 15°C to 20°C, 15°C to 25°C It is cooled to 15°C-30°C, 15°C-40°C, 15°C-50°C, 20°C-25°C, 20°C-30°C, 20°C-40°C, 20°C-50°C, 25°C-30°C, 25°C-40°C, 25°C-60°C, 25°C-80°C, 30°C-40°C, 30°C-50°C, 30°C-80°C, 50°C-80°C, 50°C-100°C, 70°C-80°C, 70°C-100°C, or 80°C-100°C. In some examples, the base plate or multiple components shrink by approximately 1%-50%. In some examples, the base plate or multiple components shrink by approximately 1%-25%. In some examples, the base plate or multiple components shrink by 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 0.8%, 1%, 1.2%, 1.5%, 1.8%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, or 50%.In some examples, the base plate or multiple components shrink by at least approximately 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 0.8%, 1%, 1.2%, 1.5%, 1.8%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, or 50%. In some examples, the base plate or multiple components may shrink by up to approximately 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 0.8%, 1%, 1.2%, 1.5%, 1.8%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, or 50%.In some examples, the base plate or multiple components are 0.01%~0.05%, 0.01%~0.1%, 0.01%~0.2%, 0.01%~0.5%, 0.05%~0.1%, 0.05%~0.2%, 0.05%~0.5%, 0.05%~0.8%, 0.1%~0.2%, 0.1%~0.5%, 0.1%~0.8%, 0.1%~1%, 0.1%~1.2%, 0.1%~1.5%, 0.2%~0.5%, 0.2%~0.8%, 0.2%~1%, 0.2%~1.2% %, 0.2%~1.5%, 0.5%~0.8%, 0.5%~1%, 0.5%~1.2%, 0.5%~1.5%, 0.5%~1.8%, 0.5%~2%, 0.5%~2.5%, 0.8%~1%, 0.8%~1.2%, 0.8%~1.5%, 0.8%~1.8%, 0.8%~2%, 0.8%~2.5%, 1%~1.2%, 1%~1.5%, 1%~1.8%, 1%~2%, 1%~2.5%, 1.2%~1.5%, 1.2%~1.8%, 1.2%~2%, 1.2%~2.5%, 1.5%~1.8%, 1.5%~2%, 1.5%~2.5%, 1.8%~2%, 1.8%~2.5%, 2%~2.5%, 2%~3%, 2%~4%, 2%~5%, 2%~8%, 2%~10%, 3%~4%, 3%~5%, 3%~8%, 3%~10%, 3%~15%, 3%~20%, 3%~25%, 4%~5%, 4%~8%, 4%~10%, 4%~15%, 4%~20%, 5%~8%, 5%~10%, 5%~15%, 5%~20%, 5%~25%, 5%~30%, 5%~50 It shrinks by %, 8%~10%, 8%~15%, 8%~20%, 8%~25%, 8%~50%, 10%~15%, 10%~20%, 10%~25%, 10%~30%, 10%~40%, 10%~50%, 15%~20%, 15%~25%, 15%~30%, 15%~40%, 15%~50%, 20%~25%, 20%~30%, 20%~40%, 20%~50%, 25%~30%, 25%~40%, 25%~50%, 30%~40%, 30%~50%, or 40%~50%.

[0172]

[0217] In some examples, one or more operations include heating a cover plate. The cover plate may be a metal cover plate, such as stainless steel, as described herein. In some examples, heating the cover plate causes it to expand, as shown, for example, in Figure 17. The cover plate may be heated to a predetermined amount, for example, an amount sufficient to cover a number of compartments arranged on the base plate. In some examples, the cover plate is heated to about 0°C to about 250°C. In some examples, the cover plate is heated to about 20°C to about 100°C. In some examples, the cover plate is heated to about 0°C, 10°C, 20°C, 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C, 100°C, 120°C, 150°C, 180°C, 200°C, 220°C, or 250°C. In some examples, the cover plate is heated to at least approximately 0°C, 10°C, 20°C, 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C, 100°C, 120°C, 150°C, 180°C, 200°C, 220°C, or 250°C. In some examples, the cover plate is heated to a maximum of approximately 0°C, 10°C, 20°C, 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C, 100°C, 120°C, 150°C, 180°C, 200°C, 220°C, or 250°C.In some cases, the cover plate is suitable for temperatures of approximately 0°C to 10°C, 0°C to 20°C, 0°C to 30°C, 0°C to 40°C, 0°C to 50°C, 0°C to 70°C, 0°C to 90°C, 10°C to 20°C, 10°C to 30°C, 10°C to 40°C, 10°C to 50°C, 10°C to 80°C, 10°C to 100°C, 20°C to 30°C, 20°C to 40°C, 20°C to 50°C, and 2 0℃~80℃, 20℃~100℃, 30℃~40℃, 30℃~50℃, 30℃~70℃, 30℃~90℃, 40℃~50℃, 40℃~60℃, 40℃~80℃, 40℃~100℃, 50℃~60℃, 50℃~80℃, 50℃~100℃, 60℃~70℃, 60℃~90℃, 60℃~120℃, 70℃~80℃, 70℃~ 90℃, 70℃~120℃, 80℃~100℃, 80℃~120℃, 80℃~150℃, 90℃~100℃, 90℃~120℃, 90℃~150℃, 100℃~120℃, 100℃~150℃, 100℃~180℃, 100℃~200℃, 100℃~220℃, 100℃~250℃, 120℃~150℃, 120℃~ It is heated to 180°C, 120°C-200°C, 120°C-220°C, 120°C-250°C, 150°C-180°C, 150°C-200°C, 150°C-220°C, 150°C-250°C, 180°C-200°C, 180°C-220°C, 180°C-250°C, 200°C-220°C, 200°C-250°C, or 220°C-250°C. In some cases, the cover plate expands by approximately 1-50%. In some cases, the cover plate expands by approximately 1-25%. In some cases, the cover plate expands by 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 0.8%, 1%, 1.2%, 1.5%, 1.8%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, or 50%. In some cases, the cover plate expands by at least 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 0.8%, 1%, 1.2%, 1.5%, 1.8%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, or 50%.In some examples, the cover plate expands by up to 0.01%, 0.05%, 0.1%, 0.2%, 0.5%, 0.8%, 1%, 1.2%, 1.5%, 1.8%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 40%, or 50%. In some examples, the cover plate expands by 0.01% - 0.05%, 0.01% - 0.1%, 0.01% - 0.2%, 0.01% - 0.5%, 0.05% - 0.1%, 0.05% - 0.2%, 0.05% - 0.5%, 0.05% - 0.8%, 0.1% - 0.2%, 0.1% - 0.5%, 0.1% - 0.8%, 0.1% - 1%, 0.1% - 1.2%, 0.1% - 1.5%, 0.2% - 0.5%, 0.2% - 0.8%, 0.2% - 1%, 0.2% - 1.2%, 0.2% - 1.5%, 0.5% - 0.8%, 0.5% - 1%, 0.5% - 1.2%, 0.5% - 1.5%, 0.5% - 1.8%, 0.5% - 2%, 0.5% - 2.5%, 0.8% - 1%, 0.8% - 1.2%, 0.8% - 1.5%, 0.8% - 1.8%, 0.8% - 2%, 0.8% - 2.5%, 1% - 1.2%, 1% - 1.5%, 1% - 1.8%, 1% - 2%, 1% - 2.5%, 1.2% - 1.5%, 1.2% - 1.8%, 1.2% - 2%, 1.2% - 2.5%, 1.5% - 1.8%, 1.5% - 2%, 1.5% - 2.5%, 1.8% - 2%, 1.8% - 2.5%, 2% - 2.5%, 2% - 3%, 2% - 4%, 2% - 5%, 2% - 8%, 2% - 10%, 3% - 4%, 3% - 5%, 3% - 8%, 3% - 10%, 3% - 15%, 3% - 20%, 3% - 25%, 4% - 5%, 4% - 8%, 4% - 10%, 4% - 15%, 4% - 20%, 5% - 8%, 5% - 10%, 5% - 15%, 5% - 20%, 5% - 25%, 5% - 30%, 5% - 50%, 8% - 10%, 8% - 15%, 8% - 20%, 8% - 25%, 8% - 50%, 10% - 15%, 10% - 20%, 10% - 25%, 10% - 30%, 10% - 40%, 10% - 50%, 15% - 20%, 15% - 25%, 15% - 30%, 15% - 40%, 15% - 50%, 20% - 25%, 20% - 30%, 20% - 40%, 20% - 50%, 25% - 30%, 25% - 40%, 25% - 50%, 30% - 40%, 30% - 50%, or 40% - 50%.

[0173]

[0218] When the temperature of the cover plate, the base plate (and / or compartments), or both, is equalized, heating of the cover plate and / or cooling of the base plate (and / or compartments) can be stopped. The cover plate can then be placed or pressed down on the plurality of compartments 2020. In some examples, it may not be necessary to cool the base plate or the plurality of compartments in response to the expansion of the cover plate. The cover plate, the base plate, and / or the plurality of compartments can be returned to a predetermined temperature 2025. The predetermined temperature may be a storage temperature, such as below ambient temperature. The cover plate can shrink or contract around the plurality of compartments (e.g., thermally contract), thereby sealing the compartments and the contents therein (e.g., polynucleotides or DNA).

[0174]

[0219] For example, in the operation of FIG. 20, a temperature gradient generated across one or more components of the device or assembly can be monitored. The temperature can be monitored radiatively, inductively, or resistively. In some examples, the temperature gradient is monitored by a sensor for measuring one or more of temperature, radiation, or resistance, such as those described herein. In some examples, the sensor is part of a system that includes the device or is part of one or more compartments of the device, such as a base plate, a cover plate, or a compartment or structure described herein.

[0175]

[0220] Polynucleotides or portions thereof can be obtained from the devices described herein. Thus, in some examples, the method further includes obtaining polynucleotides or portions thereof. In some examples, obtaining includes removing a cover plate in contact with a plurality of containers, a base plate, or both. In some examples, obtaining includes heating the cover plate or a portion thereof, cooling the base plate or a portion thereof, or both. In some examples, heating the cover plate or a portion thereof causes the cover plate to expand, and / or cooling the base plate or a portion thereof causes the base plate to contract, as a result the cover plate can be removed from the plurality of compartments.

[0176]

[0221] In some examples, multiple polynucleotides or portions thereof can be obtained without removing the cover plate in contact with multiple compartments and / or the base plate. In some examples, obtaining involves perforating a portion of the cover plate. The cover plate can be perforated, for example, using a needle or syringe, or any other suitable instrument known in the art. In some examples, if the polynucleotides are stored as a solid, an instrument such as a needle or syringe may be used to dissolve multiple polynucleotides or portions thereof in solution before obtaining them from the compartments.

[0177]

[0222] The selection of dimensions or materials for the encapsulation devices described herein may involve one or more considerations. In some examples, the selection of dimensions for the cover plate, base plate (e.g., cooling plate), or compartment (e.g., glass vial) creates a strong compression seal between the cover plate and the outer edge of the compartment (e.g., the side of the glass vial). In some examples, this allows for hermetically sealing of the contents of the compartment. In some cases, when the heating or cooling process, or the sealing process, is carried out in an inert atmosphere, the polynucleotides or DNA in the compartment are protected from degradation, for example, by impurities being sealed into the final structure or by the intrusion of external impurities over time. In some examples, the integrity of the compression seal can be maintained over a relatively wide temperature range by selecting the cover plate material and compartment material (e.g., glass vial) such that they have similar temperature coefficients of expansion (TCE). In some examples, the DNA can be accessed by perforating the top of the cover plate by selecting the cover plate material and / or specifically the thickness of the cover plate above the top of the compartment. In some cases, aliquots for rehydration and / or subsequent processing (e.g., amplification, sequencing) can be enabled by puncturing a portion of the top of the cover plate. In some cases, the accessed DNA can be decoded, and the information encoded in the DNA can be recovered.

[0178]

[0223] The dimensions and materials for encapsulation described herein may illustrate exemplary embodiments. In some examples, the dimensions and materials for encapsulation described herein facilitate integration with standard biotechnology components and systems known in the art. In some examples, the dimensions and materials for encapsulation described herein illustrate a method and / or demonstrate functionality. However, the selected materials and / or dimensions shown in exemplary embodiments may not preclude the use of other materials and / or dimensions.

[0179]

[0224] The operation of successful embodiments of the encapsulation systems, methods, devices, or assemblies described herein may depend on several factors. In some examples, factors include tolerances for the dimensions of one or more components (e.g., a base plate, a cover plate, or a compartment), the materials selected for these components, the assembly of the system (e.g., used to position, heat, cool, move, and position the individual components), or any combination thereof. In some examples, the tolerances for the wall thickness of a compartment (e.g., a vial) and the 3-sigma tolerance of the external dimensions of the compartment are within a combination of tolerances for concave features of the cover plate (e.g., tapped holes) and control of the upper temperature limit to which the plate is heated. In some examples, some movement of the vial is possible when positioning the heated cover plate (e.g., considering the elasticity and flexible positioning of the components), but the cover plate can be precisely aligned, and therefore a positioning feedback system may be used when pressing the cover plate onto the vial. In some examples, the material, tolerances, or dimensions may be selected so that the gap between the concave features of the coverplate (e.g., tapped holes) and the outer edge of the compartment (e.g., vial) is small (e.g., a few microns to a few microns). In such examples, premature cooling and compression of the coverplate can be avoided. In some examples, harmful heating of the dried DNA can be avoided by limiting the maximum temperature at which the coverplate is heated. In some further examples, harmful heating of the dried DNA can be avoided by reducing its emissivity as much as possible. In some examples, the compartment contains borosilicate glass with high thermal conductivity. In some examples, the baseplate and / or compartment may be cooled to a temperature that matches the temperature at which cracks in the glass vial (e.g., due to shrinkage differences) are eliminated when the coverplate is positioned.

[0180]

[0225] In some examples, one or more considerations discussed herein are addressed by the choice of materials used to manufacture one or more components, and / or the dimensions and number of components used. In some examples, the number of components includes the number of components per unit cell. For example, due to the expansion of the cover plate when heated, a larger number of compartments (e.g., vials) per cover plate can result in a larger offset between the compartment position and the concave features (e.g., tapped holes) on the cover plate during heating and expansion. In some examples, for the periodicity of six vials, this angle may be small (e.g., about 0.45°), and larger cover plates can result in a larger offset in the alignment between the base plate / cooling plate and the cover plate. In some examples, the exemplary embodiments illustrated herein can be adjusted, for example, by higher cover plate heating (e.g., by shielding and modifying the geometric shape of the compartments), adjustment of the size and shape of the compartments, modification of the geometric shape of the cooling plate, or for different orientations or even different numbers and pitches of concave features (e.g., wells) and enclosed compartments (e.g., vials).

[0181]

[0226] Indexing of contents

[0227] Provided herein are systems and methods for indexing content for DNA data storage. A system for storing digital information may include a plurality of polynucleotides. The plurality of polynucleotides may collectively encode digital information. A system for storing digital information may further include a structure for storing the plurality of polynucleotides. The structure may include means for indexing based on the content stored within the structure. In some examples, the means for indexing include tags, such as radio frequency identification (RFID) tags. The tags may include metadata related to the content, such as the plurality of polynucleotides or digital information stored in the structure. In some examples, the tags may further include an association between the digital information encoded in the plurality of polynucleotides and an external file system or database. In some examples, the tags may be labels, markers, identifiers, or any variation thereof. The tags may be part of a structure for polynucleotide storage, such as those provided herein.

[0182]

[0228] A tag may contain one or more feature parts. These feature parts may enable the tag to function as a label, file system, database, or a combination thereof. In some examples, the file system is a dynamic file system. In some cases, one or more features include remote accessibility. In some examples, the entire contents or parts of the contents of one or more structures for DNA data storage are cataloged remotely within the storage system and / or after the structures are removed from the storage system. In some cases, one or more feature parts include identification and / or selection of individual structures. In some examples, individual structures are identified and / or selected from multiple structures based on the tag for sequencing. In some cases, one or more feature parts include knowledge of the arrangement of all or part of the structures within the storage system. In some examples, the arrangement of structures within the storage system can be known in real time by system control software. In some examples, the arrangement of structures is presented to a system host application. In some cases, one or more feature parts include the ability to update the tag. In some examples, after the contents of a structure have been copied (e.g., PCR) and sequenced to access digital information, the tag may be updated to reflect access to the contents within the structure. In some cases, one or more feature parts include security. In some examples, tags are incorporated into the structure so that the origin of multiple polynucleotides in the capsule is undeniable. For example, if the contents within the structure are accessed, the tags are modified. In some cases, one or more feature parts include fixation. In some examples, for the purpose of fixation checking, tags provide physically directly associated information about all or some aspects of the contents of the structure (e.g., synthesis date, subsequent processing steps, volume label, and number of associated copies).

[0183]

[0229] Tags can be used to identify and / or retrieve content stored in a structure (e.g., digital information). In some examples, tags include barcodes. In some examples, barcodes include optical barcodes. In some examples, barcodes include linear barcodes or matrix (2D) barcodes. In some examples, tags include electromagnetic tags. Electromagnetic tags can be identified using frequencies in the electromagnetic spectrum. Frequencies in the electromagnetic spectrum may include radio frequencies, microwave frequencies, infrared frequencies, visible frequencies, UV frequencies, X-ray frequencies, or gamma-ray frequencies.

[0184]

[0230] In some examples, the tags include radio frequency identification (RFID) tags. In some examples, the RFID tags have low frequency (LF), high frequency (HF), or ultra-high frequency (UHF) RFID tags. LF-type RFID tags have a frequency range of approximately 30 kHz to 300 kHz. HF-type RFID tags have a frequency range of approximately 3 MHz to 30 MHz. In some examples, HF-type RFID tags have a frequency range of approximately 10 to 20 MHz. In some examples, HF-type RFID tags have a frequency range of approximately 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 MHz. In some examples, HF-type RFID tags have a frequency range of at least approximately 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 MHz. In some examples, HF RFID tags have a frequency range of approximately 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 MHz. In some examples, HF RFID tags have a frequency range of approximately 13.56 MHz. UHF RFID tags have a frequency range of approximately 300 MHz and 3 GHz. In some examples, UHF RFID tags have a frequency range of approximately 300 MHz to approximately 1000 MHz. In some examples, UHF RFID tags operate in the following frequency ranges: approximately 300MHz-400MHz, 300MHz-500MHz, 300MHz-600MHz, 300MHz-700MHz, 300MHz-800MHz, 300MHz-900MHz, 300MHz-1,000MHz, 400MHz-500MHz, 400MHz-600MHz, 400MHz-700MHz, 400MHz-800MHz, 400MHz-900MHz, 400MHz-1,000MHz, and 500MHz-600MHz. The frequency range of z is 500MHz~700MHz, 500MHz~800MHz, 500MHz~900MHz, 500MHz~1,000MHz, 600MHz~700MHz, 600MHz~800MHz, 600MHz~900MHz, 600MHz~1,000MHz, 700MHz~800MHz, 700MHz~900MHz, 700MHz~1,000MHz, 800MHz~900MHz, 800MHz~1,000MHz, or 900MHz~1,000MHz.In some cases, UHF RFID tags have a frequency range of approximately 300 MHz, 400 MHz, 500 MHz, 600 MHz, 700 MHz, 800 MHz, 900 MHz, or 1,000 MHz. In some cases, UHF RFID tags have a frequency range of at least approximately 300 MHz, 400 MHz, 500 MHz, 600 MHz, 700 MHz, 800 MHz, or 900 MHz. In some cases, UHF RFID tags have a frequency range of up to approximately 400 MHz, 500 MHz, 600 MHz, 700 MHz, 800 MHz, 900 MHz, or 1,000 MHz. In some cases, UHF RFID tags have a frequency range of approximately 433 MHz. In some cases, UHF RFID tags have a frequency range of approximately 860 MHz to 960 MHz.In some examples, UHF RFID tags operate in the following frequency ranges: approximately 860MHz-870MHz, 860MHz-880MHz, 860MHz-890MHz, 860MHz-900MHz, 860MHz-910MHz, 860MHz-920MHz, 860MHz-930MHz, 860MHz-940MHz, 860MHz-950MHz, 860MHz-960MHz, 870MHz-880MHz, 870MHz-890MHz, and 870MHz-900MHz. Hz, 870MHz~910MHz, 870MHz~920MHz, 870MHz~930MHz, 870MHz~940MHz, 870MHz~950MHz, 870MHz~960MHz, 880MHz~890MHz , 880MHz~900MHz, 880MHz~910MHz, 880MHz~920MHz, 880MHz~930MHz, 880MHz~940MHz, 880MHz~950MHz, 880MHz~960MHz, 89 0MHz~900MHz, 890MHz~910MHz, 890MHz~920MHz, 890MHz~930MHz, 890MHz~940MHz, 890MHz~950MHz, 890MHz~960MHz, 900M Hz~910MHz, 900MHz~920MHz, 900MHz~930MHz, 900MHz~940MHz, 900MHz~950MHz, 900MHz~960MHz, 910MHz~920MHz, 910MHz~ The frequency ranges are 930MHz, 910MHz-940MHz, 910MHz-950MHz, 910MHz-960MHz, 920MHz-930MHz, 920MHz-940MHz, 920MHz-950MHz, 920MHz-960MHz, 930MHz-940MHz, 930MHz-950MHz, 930MHz-960MHz, 940MHz-950MHz, 940MHz-960MHz, or 950MHz-960MHz. In some examples, UHF RFID tags have frequency ranges of approximately 860MHz, 870MHz, 880MHz, 890MHz, 900MHz, 910MHz, 920MHz, 930MHz, 940MHz, 950MHz, or 960MHz.In some cases, UHF RFID tags have a frequency range of at least approximately 860 MHz, 870 MHz, 880 MHz, 890 MHz, 900 MHz, 910 MHz, 920 MHz, 930 MHz, 940 MHz, or 950 MHz. In some cases, UHF RFID tags have a frequency range of up to approximately 870 MHz, 880 MHz, 890 MHz, 900 MHz, 910 MHz, 920 MHz, 930 MHz, 940 MHz, 950 MHz, or 960 MHz.

[0185]

[0231] A tag can be an active tag, a passive tag, or a semi-passive tag. An active tag generally refers to a tag that has a transmitter and a power supply. A passive tag generally refers to a power-less tag that receives a signal from a reader with an antenna, and the tag reflects the energy back to the reader. A semi-passive tag generally refers to a passive tag that has a power supply used to turn on the tag in the presence of a signal. In some examples, the tag is an RFID tag. In some examples, the RFID tag is an active RFID tag, a passive RFID tag, or a semi-passive RFID tag. In some examples, the passive tag is a UHF RFID tag. In some examples, the passive tag is a Near Field Communication (NFC) tag.

[0186]

[0232] Tags may include metadata related to the content stored in the structure. Metadata related to the stored content of the structure (e.g., multiple polynucleotides and / or digital information) may, in non-limiting examples, include data type, data size, data format, encryption codec, synthesis date, last access date, last handling date, owner information, manufacturing information, storage mechanism, or any combination thereof. Metadata may further include any other information related to the synthesis, storage, and / or any other handling processes of the multiple polynucleotides in the structure (e.g., volume, temperature, copy number, etc.). In some examples, data type includes attributes of the data (e.g., integer, boolean, real, complex, string, tuple, double, byte, list, byte, array, short, long, floating, character, vector, stack, set, frozen set, dictionary, etc.). In some examples, data size is the size of the digital information encoded as multiple polynucleotides. Data size may include, but is not limited to, the size of the information items described herein. In some examples, data format includes the format of the information items as described herein. In some examples, the encryption codec may include an error correction code (ECC). The ECC may include, in non-limiting examples, Reed-Solomon (RS) codes, LDPC codes, pole codes, turbo codes, or any combination thereof. In some examples, the synthesis date includes the synthesis date of multiple polynucleotides. In some examples, the last access date includes the last date on which a structural tag was scanned, the structure was opened, the polynucleotide was a whole or partial sequence, the polynucleotide was decrypted whole or partially, or any combination thereof. In some examples, the previously processed date includes, for example, the date on which the structure was moved or accessed by means previously described. In some examples, the owner information includes information about the owner of the digital information, multiple polynucleotides, the structure, or any combination thereof. In some examples, the manufacturer information includes information about the manufacturer of multiple polynucleotides, the structure, or any combination thereof.In some examples, the preservation mechanism includes preservation mechanisms for multiple polynucleotides, such as dehydration mechanisms, ionic solvent mechanisms, salt-based containment mechanisms, glass-based containment mechanisms, or any combination thereof.

[0187]

[0233] The size of metadata can be based at least partially on the size of the structure, the format of the metadata, the size of the tags, or any combination thereof. In some examples, the size of the metadata is at least partly based on International Organization for Standardization (ISO) standards. In some examples, the metadata is approximately 0.1 to 5 kB. In some examples, the metadata is approximately 0.1 kB to 0.5 kB, 0.1 kB to 1 kB, 0.1 kB to 1.5 kB, 0.1 kB to 2 kB, 0.1 kB to 2.5 kB, 0.1 kB to 3 kB, 0.1 kB to 3.5 kB, 0.1 kB to 4 kB, 0.1 kB to 4.5 kB, 0.1 kB to 5 kB, 0.5 kB to 1 kB, 0.5 kB to 1. 5kB, 0.5kB~2kB, 0.5kB~2.5kB, 0.5kB~3kB, 0.5kB~3.5kB, 0.5kB~4kB, 0.5kB~4.5kB, 0.5 kB~5kB, 1kB~1.5kB, 1kB~2kB, 1kB~2.5kB, 1kB~3kB, 1kB~3.5kB, 1kB~4kB, 1kB~4.5kB, 1k B~5kB, 1.5kB~2kB, 1.5kB~2.5kB, 1.5kB~3kB, 1.5kB~3.5kB, 1.5kB~4kB, 1.5kB~4.5kB, 1 .5kB~5kB, 2kB~2.5kB, 2kB~3kB, 2kB~3.5kB, 2kB~4kB, 2kB~4.5kB, 2kB~5kB, 2.5kB~3kB, The metadata ranges from 2.5kB to 3.5kB, 2.5kB to 4kB, 2.5kB to 4.5kB, 2.5kB to 5kB, 3kB to 3.5kB, 3kB to 4kB, 3kB to 4.5kB, 3kB to 5kB, 3.5kB to 4kB, 3.5kB to 4.5kB, 3.5kB to 5kB, 4kB to 4.5kB, 4kB to 5kB, or approximately 4.5kB to 5kB. In some examples, the metadata is approximately 0.1kB, approximately 0.5kB, approximately 1kB, approximately 1.5kB, approximately 2kB, approximately 2.5kB, approximately 3kB, approximately 3.5kB, approximately 4kB, approximately 4.5kB, or approximately 5kB. In some examples, the metadata is at least approximately 0.1kB, 0.5kB, 1kB, 1.5kB, 2kB, 2.5kB, 3kB, 3.5kB, 4kB, or 4.5kB. In some examples, the metadata is at most approximately 0.5kB, 1kB, 1.5kB, 2kB, 2.5kB, 3kB, 3.5kB, 4kB, 4.5kB, or 5kB.

[0188]

[0234] In some examples, two or more structures may contain the same polynucleotides. Two or more structures having the same polynucleotides may provide redundancy for stored digital information. Each of the two or more structures may contain an RFID tag. In some examples, each RFID tag contains metadata pointing to another structure containing the same polynucleotides. For example, if structures A and B contain the same polynucleotides, tag A on structure A may contain metadata such as an identification (ID) number or a universally unique identifier (UUID) for structure B. Similarly, tag B on structure B may contain metadata such as the identification information for structure A.

[0189]

[0235] Metadata on a tag can reduce the redundancy of stored digital information. In some examples, reduced redundancy increases the payload (e.g., digital information) stored in multiple polynucleotides. In some examples, reduced redundancy increases storage capacity. In some examples, metadata on a tag points to one or more other structures containing multiple polynucleotides. In some examples, the metadata on a tag provides the position of a structure relative to other structures. In some examples, the metadata on a tag provides the position of multiple polynucleotides relative to other polynucleotides in other structures. In some examples, the metadata indicates the order of structures. Referring to Figure 14, one or more digital information items 1410 can be divided into multiple sub-items 1420. Multiple sub-items 1420 can be coded as multiple polynucleotides, each of which is stored in a corresponding structure 1430. In such examples, the metadata for each of the structures 1430 includes the order of the structures (e.g., 1, 2, 3, 4, etc.) and can be used to reconstruct one or more information items.

[0190]

[0236] Structures within a data storage system can be indexed using tags. In some examples, a method for storing digital information includes one or more of the following: (a) synthesizing multiple polynucleotides that collectively encode the digital information; (b) writing metadata about the multiple polynucleotides to a tag; and (c) storing the multiple polynucleotides within a structure. In some examples, the tag is an RFID tag. In some examples, the structure includes an RFID tag. In some examples, multiple polynucleotides are synthesized in (a) and metadata about the multiple polynucleotides is written simultaneously in (b). In some examples, multiple polynucleotides are synthesized in (a) and metadata about the multiple polynucleotides is written sequentially in (b). In some examples, one or more of (a), (b), and (c) are performed autonomously.

[0191]

[0237] Figure 11 provides an exemplary method for storing digital information in multiple polynucleotides using tags for content indexing. Digital information, such as items of information described herein, can be coded as multiple polynucleotides.1110 In some examples, the digital information is coded as multiple polynucleotides using a codec. In some examples, the codec is a high-level codec, a low-level codec, or a combination thereof, such as those described herein. In some examples, the codec includes an error-correcting code (ECC), such as those described herein. Multiple polynucleotides that collectively code the digital information are then synthesized (e.g., in a synthesizer unit).1115 In some examples, the multiple polynucleotides are synthesized using ligation-based synthesis, enzyme-based synthesis, or phosphoramidite-based synthesis. In some examples, the multiple polynucleotides are stored using synthesis methods further provided herein. Metadata related to the multiple polynucleotides, such as those provided herein, is written to an RFID tag.1120 In some examples, the RFID tag is a UHF RFID tag. In some examples, the RFID tag is a passive tag. Then, multiple polynucleotides are stored in the structure containing the RFID tag. In some examples, the multiple polynucleotides are stored in the structure before the metadata is written to the RFID tag. In some examples, the multiple polynucleotides are stored in the structure after the metadata has been written to the RFID tag. The structure can be stored in a DNA data storage system. In some examples, the DNA data storage system is partially autonomous. In some examples, the DNA data storage system is fully autonomous.

[0192]

[0238] Figure 12 provides an exemplary method for retrieving digital information within multiple polynucleotides using tags for content indexing. Digital information, such as items of information described herein, may be encoded as multiple polynucleotides and stored in a structure containing tags in a DNA data storage system. In some examples, the tags are RFID tags. The RFID tags can be read by a reader (e.g., scanned 1210). In some examples, the reader scans multiple structures containing RFID tags in parallel (e.g., 100 or 1000 structures). In some examples, the RFID tags are remotely accessed. In some examples, a structure storing multiple polynucleotides encoding the desired information may be identified based on metadata stored in the RFID tags. The multiple polynucleotides are then retrieved 1215. The retrieved polynucleotides may be sequenced (e.g., in a sequencer unit) to provide a digital output containing the sequences of the multiple polynucleotides 1220. In some examples, the multiple polynucleotides are amplified in an amplification chamber before sequencing. The sequences of the multiple polynucleotides are decoded to retrieve the digital information 1225. In some cases, the sequence is decoded using a codec. In some cases, the codec is a high-level codec, a low-level codec, or a combination thereof, such as those described herein. In some cases, the codec includes an error correction code (ECC), such as those described herein. In some cases, once multiple polynucleotides have been accessed, the RFID tag is updated to reflect the access.

[0193]

[0239] Referring to Figure 3, an exemplary schematic diagram of a passive RFID system is provided. In a passive RFID system, the passive RFID tag 1310 includes an integrated circuit or chip for storing and processing information. The integrated circuit or chip can further modulate and / or demodulate radio frequency (RF) signals. The RFID tag may include a tag antenna 1315 capable of receiving RF signals. Upon receiving an RF signal, an electric and magnetic field is generated. The RFID tag 1310 can draw power from the electric and magnetic fields of the integrated circuit. RF signals can be transmitted using an RFID reader 1320, which includes a reader antenna 1325. In some examples, the RFID reader 1320 is powered. The integrated circuit or chip of the RFID tag 1310 can modulate the backscattered RF signal and send it back to the RFID reader 1320. The backscattered RF signal may contain information (e.g., metadata) encoded in the memory of the RFID tag 1310. In some examples, the memory is non-volatile memory. In some examples, the memory is rewritable.

[0194]

[0240] An RFID reader can scan one or more RFID tags in parallel. In some examples, an RFID reader can scan approximately 10 to 5000 RFID tags in parallel. In some examples, an RFID reader can scan approximately 10 to 50, 10 to 100, 10 to 150, 10 to 200, 10 to 250, 10 to 500, 10 to 1,000, 10 to 1,500, 10 to 2,000, 10 to 2,500, 10 to 5,000, 50 to 100, 50 to 150, 50 to 200, 50 to 250, 50 to 500, 50 to 1,000, 50 to 1,500, 50 to 2, 000, 50~2,500, 50~5,000, 100~150, 100~200, 100~250, 100~500, 100~1,000, 100~1,500, 100~2,000, 100~2,500, 100~5,000, 150~200, 150~250, 150~500, 150~1,000, 150~1,500, 150~2,000, 150~2 500, 150~5,000, 200~250, 200~500, 200~1,000, 200~1,500, 200~2,000, 200~2,500, 200~5,000, 250~500, 250~1,000, 250~1,500, 250~2,000, 250~2,500, 250~5,000, 500~1,000, 500~1,500, 500~2, 000, 500-2,500, 500-5,000, 1,000-1,500, 1,000-2,000, 1,000-2,500, 1,000-5,000, 1,500-2,000, 1,500-2,500, 1,500-5,000, 2,000-2,500, 2,000-5,000, or 2,500-5,000 RFID tags are scanned in parallel. In some examples, the RFID reader scans approximately 10, 50, 100, 150, 200, 250, 500, 1,000, 1,500, 2,000, 2,500, or 5,000 RFID tags in parallel. In some cases, an RFID reader can scan up to approximately 10, 50, 100, 150, 200, 250, 500, 1000, 1500, 2000, 2500, or 5000 RFID tags in parallel.In some examples, the RFID reader scans at least about 10, 50, 100, 150, 200, 250, 500, 1000, 1500, 2000, 2500, or 5000 RFID tags in parallel.

[0195]

[0241] The RFID reader can scan one or more RFID tags from a remote location. In some examples, the distance is from about 0.1 meter (m) to about 6 m. In some examples, the distance is 0.1 m to 0.25 m, 0.1 m to 0.5 m, 0.1 m to 0.75 m, 0.1 m to 1 m, 0.1 m to 1.5 m, 0.1 m to 2 m, 0.1 m to 2.5 m, 0.1 m to 3 m, 0.1 m to 4 m, 0.1 m to 5 m, 0.1 m to 6 m, 0.25 m to 0.5 m, 0.25 m to 0.75 m, 0.25 m to 1 m, 0.25 m to 1.5 m, 0.25 m to 2 m, 0.25 m to 2.5 m, 0.25 m to 3 m, 0.25 m to 4 m, 0.25 m to 5 m, 0.25 m to 6 m, 0.5 m to 0.75 m, 0.5 m to 1 m, 0.5 m to 1.5 m, 0.5 m to 2 m, 0.5 m to 2.5 m, 0.5 m to 3 m, 0.5 m to 4 m, 0.5 m to 5 m, 0.5 m to 6 m, 0.75 m to 1 m, 0.75 m to 1.5 m, 0.75 m to 2 m, 0.75 m to 2.5 m, 0.75 m to 3 m, 0.75 m to 4 m, 0.75 m to 5 m, 0.75 m to 6 m, 1 m to 1.5 m, 1 m to 2 m, 1 m to 2.5 m, 1 m to 3 m, 1 m to 4 m, 1 m to 5 m, 1 m to 6 m, 1.5 m to 2 m, 1.5 m to 2.5 m, 1.5 m to 3 m, 1.5 m to 4 m, 1.5 m to 5 m, 1.5 m to 6 m, 2 m to 2.5 m, 2 m to 3 m, 2 m to 4 m, 2 m to 5 m, 2 m to 6 m, 2.5 m to 3 m, 2.5 m to 4 m, 2.5 m to 5 m, 2.5 m to 6 m, 3 m to 4 m, 3 m to 5 m, 3 m to 6 m, 4 m to 5 m, 4 m to 6 m, or 5 m to 6 m. In some examples, the distance is 0.1 m, 0.25 m, 0.5 m, 0.75 m, 1 m, 1.5 m, 2 m, 2.5 m, 3 m, 4 m, 5 m, or 6 m. In some examples, the distance is at least 0.1 m, 0.25 m, 0.5 m, 0.75 m, 1 m, 1.5 m, 2 m, 2.5 m, 3 m, 4 m, or 5 m. In some examples, the distance is at most 0.25 m, 0.5 m, 0.75 m, 1 m, 1.5 m, 2 m, 2.5 m, 3 m, 4 m, 5 m, or 6 m.

[0196]

[0242] The methods for indexing content described herein may be used to determine the data integrity of stored digital information. The digital information may be stored in a structure including a tag, as described herein. A method for determining the data integrity of stored digital information may include one or more of the following: (a) writing metadata about a plurality of polynucleotides to an RFID tag; and (b) scanning the RFID tag after a period of time to determine the data integrity. In some examples, the method further includes one or more of the following: (c) providing a plurality of polynucleotides to be collectively coded for the digital information; and (d) storing the plurality of polynucleotides in a structure. In some examples, the structure includes an RFID tag.

[0197]

[0243] RFID tags can be enabled or disabled during scanning. In some examples, a valid scan of an RFID tag is error-free. Without errors, a backscattered RF signal is transmitted from the RFID tag to the RFID reader. The backscattered RF signal may contain metadata encoded in the RFID tag's memory. The metadata may be related to multiple polynucleotides stored in the structure containing the RFID tag. In some examples, a valid scan of an RFID tag is validated by the reader based at least partially on the metadata received. In some examples, the metadata encoded in the RFID tag is matched against information in a database or file system. In some examples, the RFID tag includes a Trusted Platform Module (TPM). The TPM may also use systems and methods to ensure the integrity of the RFID tag, such as those provided in Mubarak et al., Mutual Attestation Using TPM for Trusted RFID Protocol, Second International Conference on Network Applications, Protocols and Services, 2010, pp. 153-158. In some examples, an invalid scan of an RFID tag contains errors. In some cases, an RFID tag is invalid if it is not scannable or readable. An RFID tag may not be scannable if the RFID tag and / or the structure containing the RFID tag are damaged. Non-limiting examples of an invalid RFID tag may include the structure being exposed to corrosive materials, pulverized materials, water, heat, humidity, or any combination thereof. In some cases, an RFID tag is invalid if the structure is accessed without authorization and / or tampered with. In some cases, the connection between the RFID tag and the antenna is broken if the structure is tampered with.

[0198]

[0244] RFID tags may be updated when a structure is accessed. In some cases, RFID tags are updated (or deactivated) each time the structure is accessed. RFID tags may be updated during one or more handling steps of multiple polynucleotides. One or more handling steps may include, in non-limiting examples, synthesis, storage, sequencing, amplification, or any combination thereof. RFID tags may be updated with metadata such as date, time, location, and / or any handling information associated with one or more handling steps. Updating RFID tags during access to a structure can provide data occurrence, fixation, security, or any combination thereof. If data is duplicated (e.g., during amplification) and stored in a new capsule, a new identifying RFID tag may be written. This new RFID tag can uniquely identify the contents. In some cases, the new RFID tag contains metadata about one or more process steps (e.g., date, time, temperature, duration, etc. of amplification). In some cases, the new RFID tag contains metadata related to one or more "parent" structures. In some cases, the original RFID tag and the new RFID tag have the same metadata, either entirely or partially. In some cases, the original RFID tag and the new RFID tag have different metadata.

[0199]

[0245] In some examples, the period between when an RFID tag is scanned ranges from approximately 1 year to approximately 100 years. In some examples, the periods are 1-2 years, 1-5 years, 1-10 years, 1-20 years, 1-30 years, 1-40 years, 1-50 years, 1-60 years, 1-70 years, 1-80 years, 1-90 years, 1-100 years, 2-5 years, 2-10 years, 2-20 years, 2-30 years, 2-40 years, 2-50 years, 2-60 years, 2-70 years, and 2 years. ~80 years, 2 years ~ 90 years, 2 years ~ 100 years, 5 years ~ 10 years, 5 years ~ 20 years, 5 years ~ 30 years, 5 years ~ 40 years, 5 years ~ 50 years, 5 years ~ 60 years, 5 years ~ 70 years, 5 years ~ 80 years, 5 ~90 years, 5~100 years, 10~20 years, 10~30 years, 10~40 years, 10~50 years, 10~60 years, 10~70 years, 10~80 years, 10~90 years , 10 to 100 years, 20 to 30 years, 20 to 40 years, 20 to 50 years, 20 to 60 years, 20 to 70 years, 20 to 80 years, 20 to 90 years, 20 to 90 years, 30 years ~40 years, 30 to 50 years, 30 to 60 years, 30 to 70 years, 30 to 80 years, 30 to 90 years, 30 to 100 years, 40 to 50 years, 40 to 60 years, 40 to 70 years, The periods are 40-80 years, 40-90 years, 40-100 years, 50-60 years, 50-70 years, 50-80 years, 50-90 years, 50-100 years, 60-70 years, 60-80 years, 60-90 years, 60-100 years, 70-80 years, 70-90 years, 70-100 years, 80-90 years, 80-100 years, or 90-100 years. In some examples, the periods are 1 year, 2 years, 5 years, 10 years, 20 years, 30 years, 40 years, 50 years, 60 years, 70 years, 80 years, 90 years, or 100 years. In some examples, the duration is at least 1, 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, or 90 years. In some examples, the duration is at most 2, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 years.

[0200]

[0246] During such storage periods, the minimum amount of digital information stored in polynucleotides is lost using the methods, systems, and devices described herein (e.g., unrecoverable after acquisition, sequencing, and decoding). Losses include the loss of bases, the loss of a sequence of bases, or the loss of digital information units (such as bits, bytes, trites, or other units). For example, after 5 years of storage at 25°C, less than 0.1%, 0.5%, 1%, 2%, or 5% of the digital information is lost. In some cases, after 5 years of storage at 30°C, less than 0.1%, 0.5%, 1%, 2%, or 5% of the digital information is lost. In some cases, after 5 years of storage at 35°C, less than 0.1%, 0.5%, 1%, 2%, or 5% of the digital information is lost.

[0201]

[0247] Post-processing

[0248] One or more components, such as a flow cell, chamber, or other components, may be used for post-processing. In some examples, post-processing includes the addition, purification, or other steps performed after synthesis and extraction of the adapter. In some examples, post-processing operations are performed after storage. A system for storing polynucleotides may further include amplification chambers 1820. Amplification units may be used to amplify multiple polynucleotides. In some examples, the system includes two or more amplification chambers 1820. In some examples, a structure is selected from a storage unit 1815, and polynucleotides derived from the structure are transferred to the amplification chambers 1820. In some examples, polynucleotides from a synthesizer unit 1810 are transferred to the amplification chambers 1820 for size selection, PCR, or other types of amplification or preparation for storage. Size selection generally involves selecting DNA of a target size and blocking much shorter or much longer strands. In some examples, filters are adjusted to capture DNA within a specific size range. In some examples, alternative methods include PCR, electrophoresis, capture with solid-phase primers complementary to the terminal sequence of the synthetic oligonucleotide, or the use of isothermal polymerase. Fluidic and / or electronic control of polynucleotide synthesis in the amplification chamber 1820 may be performed by the controller 1835. In some examples, the electronics within the amplification chamber 1820 communicate with the controller 1835. In some examples, the post-processing module and its operation are shown in Figure 2B. In some examples, the module for amplification includes one or more of the following: a thermal cycler, a seal, and chambers for rehydration and aliquot splitting.

[0202]

[0249] Coding

[0250] This specification provides devices, assemblies, compositions, systems, and methods for nucleic acid-based information (data) storage. In some examples, the devices, compositions, platforms, systems, or methods provided herein are used for DNA data storage of biomolecules extracted from a substrate. In some examples, these devices, assemblies, compositions, systems, and methods are used for encapsulating polynucleotides encoding digital information for DNA data storage. In some examples, the system comprises a base plate (and / or cooling plate), a plurality of compartments, and a cover plate. The plurality of compartments and / or the base plate may contain a material having a smaller coefficient of thermal expansion (CTE) than the material of the cover plate, so that the cover plate (which may be previously heated) can be placed on the compartments and then cooled to seal the compartments. In some further examples, these devices, compositions, systems, and methods are used for DNA data storage using radio frequency identification (RFID) tags. Biomolecules such as DNA molecules, in contrast to conventional binary information coding, provide a suitable host for storing information such as digital information, partly due to their temporal stability and enhanced information coding. In addition, biomolecules such as DNA molecules can provide high volume storage density. In the first step, a digital sequence is received that encodes an item of information (for example, digital information in binary code for computer processing). The digital sequence may contain a first set of symbols, such as binary, octal, decimal, or hexadecimal data. An encryption scheme is applied to convert the digital sequence from a first symbol string to a second symbol string. The second symbol string may contain an alternative representation of the first symbol string. In some examples, the second symbol string contains a nucleic acid sequence.

[0203]

[0251] Once the information items are converted into nucleic acid sequences, nucleic acids can be synthesized. A surface material for nucleic acid elongation, a design for gene loci (also known as arrangement spots) for nucleic acid elongation, and reagents for nucleic acid synthesis can be selected. The surface of the structure is prepared for nucleic acid synthesis. De novo polynucleotide synthesis is then carried out. The synthesized polynucleotides can be extracted whole or partially using the systems, devices, methods, or platforms provided herein. The synthesized polynucleotides are stored in the structure, and in some cases, whole or in part, are available for later release. The synthesized polynucleotides can be stored in a structure suitable for long-term storage (e.g., weeks, months, years, etc.). Structures suitable for long-term storage may be identifiable and / or cataloged, for example, using tags (e.g., barcodes or RFID tags). Upon release, all or part of the polynucleotides are sequenced and subjected to decoding to convert the nucleic acid sequence back into a digital sequence. This digital sequence is then assembled to obtain an alignment that codes the original information items.

[0204]

[0252] Information items

[0253] Optionally, the initial steps of a data storage process disclosed herein include acquiring or receiving one or more information items in the form of an initial code. In some examples, the information items are coded as a plurality of polynucleotides extracted from a substrate using a system, method, platform, or device provided herein. In some examples, the information items are coded as a plurality of polynucleotides and encapsulated using systems and methods described herein. In some examples, the system comprises a base plate (and / or cooling plate), a plurality of compartments, and a cover plate. The plurality of compartments and / or the base plate may contain a material having a smaller coefficient of thermal expansion (CTE) than the material of the cover plate, so that the cover plate (which may be previously heated) can be placed on the compartments and then cooled to seal the compartments. In some further examples, the information items are coded as a plurality of polynucleotides and stored in a structure including an RFID tag. Information items (e.g., digital information) include, without limitation, text, auditory, and visual information. Exemplary sources of information items include, without limitation, books, periodicals, electronic databases, medical records, letters, paper, audio recordings, animal records, biological profiles, broadcast programs, movies, short videos, emails, bookkeeping, call logs, internet activity logs, drawings, paintings, printed materials, photographs, pixel images, and software code. Exemplary sources of biological profiles for information items include, without limitation, gene libraries, genomes, gene expression data, and protein activity data. Exemplary formats for information items include, without limitation, .txt, .PDF, .doc, .docx, .ppt, .pptx, .xls, .xlsx, .rtf, .jpg, .gif, .psd, .bmp, .tiff, .png, and mpeg.The size of individual files encoding digital information items, or the size of multiple files encoding information items, is not limited to a maximum of 1024 bytes (equivalent to 1 KB), 1024 KB (equivalent to 1 MB), 1024 MB (equivalent to 1 GB), 1024 GB (equivalent to 1 TB), 1024 TB (equivalent to 1 PB), 1 exabyte, 1 zettabyte, 1 yottabyte, 1 xenotabyte, or more. In some examples, the amount of digital information is at least 1 gigabyte (GB). In some examples, the amount of digital information is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000 gigabytes, or greater than 1000 gigabytes. In some examples, the amount of digital information is at least 1 terabyte (TB). In some examples, the amount of digital information is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 terabytes, or greater than 1000 terabytes. In some examples, the amount of digital information is at least 1 petabyte (PB). In some examples, the amount of digital information is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 50, 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 petabytes, or greater than 1000 petabytes. In some examples, the digital information does not include genomic data obtained from organisms. In some examples, the items of information are coded. Examples of non-restrictive coding methods include 1-bit / base, 2-bit / base, 4-bit / base, or other coding methods.

[0205]

[0254] Sequencing

[0255] Polynucleotides are extracted and / or amplified from the surface on which they are synthesized or conserved. After extraction and / or amplification of polynucleotides from the surface of the structure, the polynucleotides can be sequenced using a suitable sequencing technique. In some cases, DNA sequences present on the substrate or within structural features are read. In some cases, polynucleotides conserved on the substrate are extracted, optionally assembled into longer nucleotides, and then sequenced.

[0206]

[0256] Sequencing / Decoding

[0257] The system for storing polynucleotides may further include a sequencing unit 1825. The sequencing unit 1825 may be used to sequence multiple polynucleotides. In some examples, multiple polynucleotides are transferred from an amplification chamber 1820 to the sequencing unit 1825. In some examples, the system may include additional modules for performing additional sequencing preparation steps. In some examples, multiple polynucleotides are transferred from the amplification chamber 1820 to the sequencing unit 1825 using one or more tubes or a robotic system 1830. In some examples, the amplification chamber 1820 and the sequencing unit 1825 are fluidically coupled. Fluid and / or electronic control of polynucleotide synthesis in the sequencing unit 1825 may be performed by a controller 1835. In some examples, electronics within the sequencing unit 1825 communicate with the controller 1835.

[0207]

[0258] In some examples, the system includes large-scale sequencing of polynucleotides. In some examples, the large-scale sequencing includes high-density, highly parallel sequencers. In some examples, the system includes two or more sequencing units 1825. In some examples, the sequencing unit 1825 uses centrifugal force and / or vacuum / pressure to add or remove reagents from the sequencing unit 1825. In some examples, the sequencing unit 1825 is optical-based (e.g., light source and sensor-on-chip), nanopore-based (e.g., Oxford nanopore technology (ONT)), or includes other operations (e.g., optical-based methods such as PacBio / SMRT single-molecule smart sequencing or other sequencing technologies). In some examples, the sequencing unit 1825 uses sequencing methods provided herein. In some examples, the sequencing unit 1825 uses nanopore or other electro-sequencing technologies that benefit from bulk fluid engineering provided by semiconductor manufacturing equipment. In some examples, one or more modules described herein include cameras. A camera may be used to capture one or more optical features of polynucleotides within the module. For example, a camera may be used in a synthesizer unit, a sequencing unit, or both to capture optical features of polynucleotides bound to a surface on a solid support as described herein. In some examples, sequencing includes sequencing by the use of nanoball generation (e.g., MGI / BGI) and sequencing by synthesis. In some examples, sequencing includes sequencing by synthesis (e.g., Illumina).

[0208]

[0259] A system for storing polynucleotides may include a robotic system 1830 as described herein. The robotic system may generally be used to manipulate polynucleotides within the system. The manipulations may include, but are not limited to, movement, storage, retrieval, handling, transfer, or any combination thereof. In some examples, the robotic system transfers multiple polynucleotides between modules within the system. In some examples, the robotic system manipulates (e.g., transfers) multiple polynucleotides within a storage structure as described herein. In some examples, the robotic system manipulates (e.g., transfers) multiple polynucleotides within a rack. In some examples, the rack comprises multiple structures, each equipped with an RFID tag. In some examples, the rack includes multiple solid supports for synthesis and / or sequencing. In some examples, the robotic system includes a robotic hand or a robotic picker. In some examples, the robotic system 1830 is fully integrated with storage system control software and / or firmware in a controller 1835. In some examples, the robotic system 1830 is fully integrated with an external host application. In some examples, the robotic system 1830 is fully automated.

[0209]

[0260] A system for storing polynucleotides may include controller 1835. The controller may generally be used to control modules, components, fluid mechanics, robots, or any combination thereof. Modules, components, fluid mechanics, electronics, robots, or any combination thereof may be used for the synthesis, storage, retrieval, sequencing, and / or amplification of polynucleotides. In some examples, controller 1835 has the ability to catalog all storage structures loaded, removed, and / or stored in racks. Polynucleotides may encode the digital information described herein. Modules, components, fluid mechanics, electronics, robots, or any combination thereof may be used to perform methods, models, or algorithms such as encoding or decoding polynucleotides.

[0210]

[0261] In some examples, controller 1835 controls the physical position of multiple polynucleotides. In some examples, controller 1835 provides commands to one or more modules of the system. In some examples, controller 1835 controls a robot (e.g., robotic system 1830), actuators, and fluid valves, or any other equipment in the system. In some examples, controller 1835 enables the synchronization and control of modules for processing and / or transporting polynucleotides. In some examples, polynucleotides are processed and / or transported by hydrodynamics. In some examples, controller 1835 controls one or more valves or parameters (e.g., pressure, vacuum, temperature, volume, etc.) in the system for biomolecule extraction, as provided, for example, in Figure 16. In some examples, controller 1835 can be used to orient or adjust the orientation of flow cells in the system for biomolecule synthesis and / or extraction. This allows for system flexibility and can maximize the recovery of liquids or substances (e.g., polynucleotides). In some examples, polynucleotides are processed and / or transported via electronics. In some examples, the controller 1835 controls physical parameters in one or more modules, such as pressure, vacuum, temperature, volume (of, for example, fluid), or any combination thereof, without limitation.

[0211]

[0262] In some cases, the controller 1835 invokes an encoder module or a decoder module. In some examples, the encoder module encodes digital information as a plurality of polynucleotides. In some examples, the encoder module applies one or more codecs, such as those described herein, to the digital information. In some cases, the decoder module decodes the sequence of the plurality of polynucleotides to obtain digital information. In some examples, the decoder module applies one or more codecs, such as those described herein, to the sequence of the plurality of polynucleotides. In some examples, the decoding module performs reconstruction, error correction, and outputs digital information (e.g., binary data). In some examples, the output containing the digital information is transferred to an operating system and / or a file system. The output may be provided on a display such as a graphical user interface (GUI), or on any other suitable display, such as those described herein, for providing the digital information. In some examples, the controller 1835 is implemented on one or more software modules, such as those described herein. In some examples, the controller 1835 responds to commands from an operating system, such as those described herein.

[0212]

[0263] Encoder modules generally encode digital information as multiple polynucleotides. Encoders can apply encoding schemes to digital information. In some examples, the encoding scheme includes a codec (e.g., an internal codec) for encoding binary data as a polynucleotide sequence. In some examples, the encoding scheme includes an error correction code (ECC) (e.g., an external codec). In some examples, the need for error correction is reduced because less material is lost during material transfer by utilizing a flow cell optimized for maximum material recovery from the substrate. In some cases, the encoding scheme is designed and implemented to allow streaming read and write API access. In some cases, the encoding scheme is designed and implemented to accommodate the streaming of systems and methods for digital storage described herein.

[0213]

[0264] A coding scheme can generally include one or more operations. These operations may include one or more operations that manipulate or transform data (e.g., digital information). These operations may include, but are not limited to, splitting, shuffling, concatenating, transposing, translating, copying, labeling (e.g., using an index), or any combination thereof, of data or parts of data.

[0214]

[0265] In some examples, the external codec comprises an error correction code (ECC) or scheme, such as but not limited to Reed-Solomon (RS) code, low-density parity-check (LDPC) code, polarity code, turbo code, or any variation thereof. This external codec is used to diffuse the stored digital or binary data into many oligonucleotides. In some examples, diffusing the data increases redundancy for correcting erasure (e.g., lost oligonucleotides). In some further embodiments, diffusing the data also increases redundancy for error correction from the internal codec. In some examples, the method for encoding digital or binary data in multiple nucleotide sequences includes an internal codec. In some examples, the internal codec is applied to binary data. In some examples, the internal codec is applied to binary data from an ECC. In some examples, the internal codec is applied to a lane of binary data. In some examples, the internal codec is applied to shuffled binary data.

[0215]

[0266] In some examples, the coding scheme includes an internal codec. In some examples, the internal codec is applied to encode binary data as a polynucleotide sequence. The internal codec is used to convert digital or binary data into nucleotide bases. In some examples, the internal codec can correct deletion, substitution, or insertion errors, or any combination thereof. In some further embodiments, the internal codec is used to validate oligonucleotides and discard oligonucleotides that fail the error checking algorithm to avoid contamination of external decoding. The internal codec further encodes an index, which may enable efficient clustering during decoding. In some examples, the coding scheme adds redundancy across multiple oligonucleotide sequences. In some examples, the internal codec includes generating base candidates. In some examples, base candidates are generated using a codebook, lookup table, hash, or any suitable method known in the art. In some examples, the internal codec further includes base iteration checking. In some examples, the internal codec further includes performing GC filtering.

[0216]

[0267] A decoder module generally decodes a sequence of multiple polynucleotides to obtain digital information. The decoder can apply a decoding scheme to a sequence of multiple polynucleotides. In some examples, the decoding scheme includes an internal codec, an external codec (e.g., ECC), or a combination thereof. In some examples, the decoding scheme decodes a sequence of multiple polynucleotides to produce an output containing digital information. In some examples, the decoding scheme includes undoing an operation in the coding scheme. In some examples, the operation includes, but is not limited to, splitting, shuffling, concatenating, transposing, translating, duplicating, and labeling (e.g., using an index) data, a portion of data, or any combination thereof.

[0217]

[0268] A digital output from a sequencer unit containing sequences of multiple polynucleotides may be provided to a decoding module. In some examples, the decoder module orders, clusters, and / or aligns the sequences of multiple polynucleotides. In some examples, the decoder module includes, but is limited to, an alignment algorithm such as a pairwise alignment algorithm, a multi-sequence alignment algorithm, or any other suitable algorithm.

[0218]

[0269] In some examples, the decoding scheme includes an internal codec. In some examples, the internal codec is applied to multiple polynucleotide sequences. The internal codec is used to convert the polynucleotide sequences into digital or binary data. In some examples, the internal codec can correct deletion, substitution, or insertion errors, or any combination thereof. In some further embodiments, the internal codec is used to validate oligonucleotides and discard any suspicious oligonucleotides to avoid contamination of external decoding. In some examples, the internal codec enables efficient decoding using an index.

[0219]

[0270] An internal codec, including a decoding scheme, can be applied to multiple polynucleotide sequences. In some examples, the internal codec converts each of the multiple polynucleotide sequences into binary data. In some examples, the internal codec is applied to multiple sequenced polynucleotides. In some examples, the clustered sequences are ordered, clustered, aligned, or any combination thereof. In some examples, one or more codecs include cyclic redundancy checks (CRC).

[0220]

[0271] In some examples, the internal codec includes a greedy algorithm. A greedy algorithm generally considers only the transition from the most likely state when decoding each bit position in a sequence. In some examples, the internal codec includes a maximum likelihood (ML) algorithm. An ML algorithm generally considers the transition from all states when decoding each bit position in a sequence. In some examples, the internal codec includes a mixed greedy ML algorithm. A mixed greedy ML algorithm generally can consider the transition from multiple states when decoding each bit position in a sequence. In some examples, the internal codec features a beam search decoder or a random sampling decoder (e.g., a pure sampling decoder, a top-K sampling decoder, etc.). In some cases, a beam search decoder or random sampling decoder provides a wider range of candidate states compared to a greedy decoder. In some examples, the internal codec further includes a checksum. In some examples, the internal codec includes a hash (e.g., SHA-256). In some examples, the hash verifies that the data has been correctly decoded. In some examples, encoding and decoding are performed as streams by using a hash at the end (after ECC). In some examples, this can limit memory usage to only temporary buffers.

[0221]

[0272] In some cases, an internal codec is applied to a subset of multiple data polynucleotides. In some examples, the internal codec involves probabilistic decoding. Generally, the internal codec involves decoding polynucleotides into digital information. In some cases, the internal codec involves transforming or converting each of the subsets of multiple data polynucleotides into binary data. In some examples, the entire length of a subset of multiple data polynucleotides is converted or transformed into binary data (e.g., full decoding). In some examples, a partial length of a subset of multiple data polynucleotides is converted or transformed into binary data (e.g., partial decoding). In some examples, the partial length includes an index such as an index (e.g., lane index, frame index, UUID, content ID, etc.) as described herein. In some examples, the internal codec is applied to a subset of multiple data polynucleotides that have been sequenced. In some examples, the internal codec is applied to a subset of multiple data polynucleotides that are ordered, aligned, clustered, or any combination thereof, or none of the above.

[0222]

[0273] In some cases, multiple data polynucleotides and / or subsets of multiple data polynucleotides are encoded using the methods described herein. In some cases, multiple data polynucleotides and / or subsets of multiple data polynucleotides are decoded using the methods described herein. In some examples, the internal codec includes a greedy method. In some examples, the internal codec includes a maximum likelihood (ML) algorithm. In some examples, the internal codec includes a mixed greedy ML algorithm.

[0223]

[0274] In some cases, the probabilistic decoding of the internal codec provides the likelihood of the entire decoded sequence. In some examples, redundancy within each polynucleotide sequence helps estimate the error rate when the reference polynucleotide is unknown. For example, if the internal codec decodes the sequence with high probability and / or in very few steps, the error rate is likely to be low. As a further example, if the internal codec decodes the sequence with low probability and / or in many steps, the error rate is likely to be high.

[0224]

[0275] In some cases, data polynucleotides include indices. In some examples, indices of subsets of multiple data polynucleotides are decoded. In some examples, indices are decoded using internal codecs, external codecs, or combinations thereof, for example, but not limited to those described herein. In some cases, indices are used to estimate the relative distribution of subsets of multiple polynucleotides. In some examples, relative distribution is used to estimate the homogeneity of data polynucleotides. For example, if multiple data polynucleotides contain approximately 100,000 polynucleotide sequences and the selected subset represents 0.1% of the data polynucleotides, a distribution centered around 100 decoded indices may be expected. In some examples, changes in the relative distribution between subsets of data polynucleotides indicate a loss of homogeneity across the data polynucleotides.

[0225]

[0276] In some examples, the decoding module includes an external codec (e.g., ECC). In some examples, multiple nucleotide sequences are decoded into digital or binary data. In some examples, an external codec (e.g., ECC) is applied to the digital or binary data. In some examples, the external codec includes an ECC used to encode the data (e.g., binary data). In some examples, the ECC may be Reed-Solomon (RS) code, LDPC code, pole code, turbo code, or any combination thereof. In some examples, the decoding scheme includes soft decoding. Soft decoding generally refers to decoding by considering a range of possible values ​​(e.g., using probability estimation).

[0226]

[0277] Polynucleotides synthesized and stored on the structures described herein encode data that can be interpreted by reading the sequence of the synthesized polynucleotide and converting the sequence into computer-readable binary code. In some cases, the sequence needs to be assembled, which may be required at the nucleic acid sequencing stage or the digital sequencing stage. Polynucleotides are extracted and / or amplified from the surface on which they are synthesized or stored. After extraction and / or amplification of polynucleotides from the surface of the structure, the polynucleotides may be sequenced using a suitable sequencing technique. In some cases, DNA sequences on the substrate or within the structural features are read. In some cases, polynucleotides stored on the substrate are extracted, optionally assembled into longer nucleotides, and then sequenced.

[0227]

[0278] This specification provides a detection system comprising an apparatus having the ability to sequence stored polynucleotides either directly on a synthetic structure and / or after removal from a main structure (e.g., synthetic structure, storage structure, etc.). If the synthetic structure is an open-reel flexible material tape, the detection system comprises an apparatus for holding the structure and advancing it through a detection location, and a detector positioned close to the detection location for detecting a signal emanating from a section of the tape when that section is at the detection location. In some examples, the signal indicates the presence of polynucleotides. In some examples, the signal indicates the sequence of polynucleotides (e.g., a fluorescent signal). In some examples, the information encoded in the polynucleotides on a continuous tape is read by a computer as the tape is being transported continuously through a detector operably connected to a computer. In some examples, the detection system comprises a computer system including a polynucleotide sequencing apparatus, a database for storing and retrieving data on polynucleotide sequences, software for converting the DNA code of the polynucleotide sequences into binary code, a computer for reading the binary code, or any combination thereof.

[0228]

[0279] This specification provides sequencing systems that can be incorporated into the apparatus described herein. Various sequencing methods are well known in the art and include "base calling," which identifies the identity of bases in a target polynucleotide. In some examples, polynucleotides synthesized using the methods, apparatus, compositions, and systems described herein are sequenced after cleavage from the synthesis surface. In some examples, sequencing is performed during or concurrently with polynucleotide synthesis, where base calling is performed immediately after or immediately before the extension of nucleoside monomers into a growing polynucleotide chain. Base calling methods include measuring the current / voltage generated by base addition to the template chain by polymerase catalyst. In some examples, the synthesis surface includes an enzyme such as polymerase. In some examples, such enzyme is tethered to an electrode or to the synthesis surface. In some examples, the enzyme includes a terminal deoxynucleotidyltransferase or a variant thereof.

[0229]

[0280] Computing systems

[0281] Referring to Figure 10, a block diagram is shown illustrating an exemplary machine including a computer system 1000 (e.g., a processing or computing system) in which a set of instructions can be internally executed to cause the device to perform or execute one or more of the embodiments and / or methods for static code scheduling of the present disclosure. The components in Figure 10 are illustrative and do not limit the scope of use or functionality of any hardware, software, embedded logic components, or combinations of two or more such components that implement a particular embodiment. The computing system schematically shown in Figure 10 may be part of a data storage system, as illustrated in Figure 8.

[0230]

[0282] In various embodiments, any of the systems described herein (e.g., Figures 1-8) are operably connected to a computer and optionally automated by the computer, either locally or remotely. In various examples, the methods and systems described herein further include software programs on a computer system and their use. Thus, computerized control of the synchronization of dispensing / vacuum / replenishment functions, such as organizing and synchronizing the movement, distribution, and vacuum operations of the material depositor, is within the scope of the disclosures provided herein. In some examples, the computer system is programmed to interface between a user-specified nucleotide sequence and the location of the material depositor in order to deliver the correct building blocks and / or reagents to a specified region of the substrate (e.g., a specified locus).

[0231]

[0283] For example, a computer system, such as the system shown in Figure 10, can be used to encode data represented as a set of symbols into another set of symbols. For example, the data may be represented as numeric symbols, such as the binary values ​​of "0" and "1", and the computer system may run a program that includes a codec (e.g., error correction code such as RS code, LDPC code, Turbo code). In some examples, the computer sy...

Claims

1. A system for data storage, A computing system comprising at least one processor and instructions executable by the at least one processor for performing one or more operations, Receiving digital information, Encoding digital information in one or more nucleic acid sequences, and To synthesize a library of polynucleotides corresponding to the aforementioned nucleic acid sequence. Computing systems including, A modular rack-mount composite unit, Computer controller, One or more reservoirs, A flow cell block containing one or more flow cells, Extraction stage, Post-processing unit, Storage unit, Robot interference, and / or Rack Interface Modular rack-mount composite unit, including A system that includes this.

2. The system according to claim 1, wherein the at least one flow cell block includes at least 12 flow cells.

3. The one or more flow cells described above are One or more solid supports configured for synthesizing the polynucleotide library, and One or more ports for exchanging gas, synthesis reagents, and / or extracted polynucleotides. The system according to claim 1, including the following:

4. The modular rack-mount synthesis unit includes the extraction stage and / or the post-processing unit, and the extraction stage and / or the post-processing unit is Energy supply chain, and One or more ports for exchanging synthetic reagents and / or extracted polynucleotides. The system according to claim 1, including the following:

5. The modular rack-mount composite unit includes the storage unit, and the storage unit is Storage plate, A robot interface configured to move a storage plate, and Access Port The system according to claim 1, including the following:

6. The modular rack-mount composite unit includes the rack interface, and the rack interface is power supply, Rack reagent bulkhead, Synthesis unit reagent bulkhead, One or more reservoirs, Energy supply chain, and Pilot valve bank The system according to claim 1, including the following:

7. The system according to claim 1, wherein the storage unit provides the polynucleotide library to a sequencing unit configured to sequence the polynucleotide library.

8. It further includes one or more sensors, and the one or more sensors are The state is detected, and the state includes temperature, pressure, humidity, voltage changes, current changes, capacitance, conductivity, storage plate position, liquid volume, flow rate, and / or presence of liquid. The aforementioned state is reported to the computing system. The system according to claim 1, configured as follows.

9. The system according to claim 8, wherein the modular rack-mount synthesis unit includes the at least one flow cell block, and the one or more sensors are further configured to pause synthesis in the one or more flow cells or the at least one flow cell block.

10. The system according to claim 9, wherein the system is a first system for data storage, and the computing system is configured to redirect coded data from (i) a suspended flow cell or (ii) a suspended flow cell block to an active flow cell, an active flow cell block, or a second system for data storage.

11. The computing system further includes a cache, where digital information is held in the cache until a polynucleotide encoding the digital information is stored, according to claim 1.

12. An assembly for storing information, A plurality of compartments containing a first material, each compartment configured to receive a plurality of polynucleotides that encode information, A base plate containing the second material and A cover plate containing a third material and Includes, The coefficient of thermal expansion (CTE) of the first material, the second material, or both thereof is smaller than that of the third material. assembly.

13. The assembly according to claim 12, wherein the plurality of compartments are arranged in an array on the base plate.

14. The assembly according to claim 13, wherein the base plate, the cover plate, or both thereof include a plurality of recessed feature portions, and each portion is located at least partially within a recessed feature portion.

15. The assembly according to claim 12, wherein the first material comprises a borosilicate.

16. The assembly according to claim 12, wherein the second material has a specific heat capacity of about 0.5 J / k-°C to 2.5 J / k-°C.

17. The third material is, CTE of approximately 15 μm / m-°C to 20 μm / m-°C, Thermal conductivity of approximately 15 W / m-K to approximately 20 W / m-K, and / or Specific heat capacity of approximately 0.5 J / g-°C The assembly according to claim 12, including the assembly described in claim 12.

18. The assembly according to claim 12, wherein the third material includes stainless steel.

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