Textured surfaces for polynucleotide synthesis

A technology of polynucleotides and nucleosides, applied in the direction of combinatorial chemistry, chemical libraries, and the introduction of foreign genetic materials using vectors, etc., which can solve the problems of scalability, automation, speed, accuracy, and cost.

Pending Publication Date: 2019-08-02
TWIST BIOSCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although various methods are known for the synthesis of relatively short fragments on a small scale, these techniques are less than satisfactory in terms of scalability, automation, speed, precision and cost

Method used

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  • Textured surfaces for polynucleotide synthesis
  • Textured surfaces for polynucleotide synthesis
  • Textured surfaces for polynucleotide synthesis

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0152] Provided herein are surface preparation methods in which photolithography is followed by a reactive chemical vapor deposition (CVD) step. Optionally, an exemplary first step includes cleaning the surface of the device using the cleaning methods disclosed herein. Cleaning may include oxygen plasma treatment. In some cases, a CVD step is used to deposit a mixture having at least two molecules resulting in regions of high surface energy and regions coated with a first chemical layer that is a region of lower surface energy. The mixture may comprise molecules bound to the surface and coupled to nucleoside phosphoramidites mixed with a larger number of molecules bound to the surface but not coupled to nucleoside phosphoramidites. For the two-step dilution scheme, prior to depositing the mixture on the surface, the step involves depositing 100% of the mixture constituent molecules that are bound to the surface but not conjugated to the nucleoside phosphoramidites. The first...

Embodiment 1

[0272] Example 1: Textured Silicon Array

[0273] An array of pillars is etched into a silicon dioxide wafer to increase the surface area by a factor of 2 to 3. Figure 18-21 An image of the microfluidic device is shown. Figure 18 and 19 is a top-view image of the microfluidic device. Figure 20 is a side-view image of the microfluidic device at 52 degrees, at a scale capable of distinguishing the pillars of the textured surface. Figure 21 is a side view of a textured microfluidic device.

[0274] Textured silicon arrays were fabricated with the following characteristics: the measured height of the etched pillars was 746 nm, the width of the pillar tops was 272 nm, and the width of the pillar bottoms was 264 nm, and the thickness of the thermal oxide layer "capping" the pillars was 74nm.

[0275] Figures 22A-22B Additional images of the microfluidic device are shown. Figure 22A is a side-view image of the microfluidic device that highlights the macroscopic geometry ...

Embodiment 2

[0280] Example 2: Design, fabrication and analysis of arrays with textured loci for polynucleotide extension

[0281] array design. Silicon plates were fabricated with the following features designed for etched pillars: Array A: the height of the oxide "cap" at the top of the pillars was 122 nm; the etch depth of the pillars was 301 nm; and the pillar width at the bottom was 320 nm; and Array B: the top of the pillars The height of the oxide "cap" is 112 nm; the etch depth of the pillars is 426 nm; and the pillar width at the bottom is 316 nm.

[0282] Synthesis on fabricated boards with textured seats. Each silicon plate contains an array of clusters, each cluster having a discrete position ("locus") for nucleotide extension. Figures 23A-23B Depicted are low magnification images of locus clusters after performing polynucleotide synthesis reactions. Figure 23A Image depicting clusters of non-textured loci. Figure 23B Images depicting clusters of textured loci.

[0283]...

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Abstract

The present invention discloses methods, devices and systems \herein for surfaces for de novo polynucleotide synthesis that provide for increased polynucleotide yield. Surfaces described herein comprise a texture that increases surface area provide for increased polynucleotide yield compared to non-textured surfaces. In addition, the patterned placement of nucleoside coupling reagent spanning suchsurfaces provides for improved synthesis yield, representation, and a reduction in contamination on the surface between different polynucleotide species.

Description

[0001] cross reference [0002] This application claims the benefit of US Provisional Application No. 62 / 370,548, filed August 3, 2016, which is hereby incorporated by reference in its entirety. [0003] Incorporate by reference [0004] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. Background technique [0005] Efficient chemical gene synthesis with high fidelity and low cost plays a central role in biotechnology and medicine, as well as basic biomedical research. De novo gene synthesis is a powerful tool for basic biological research and biotechnological applications. Although various methods are known for the synthesis of relatively short fragments on a small scale, these techniques fall short in terms of scalability, automation, speed, accu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10C12N15/66C40B50/00C40B50/14C40B50/18
CPCC12N15/10C40B50/00C40B50/14C40B50/18B01J2219/00497B01J2219/00587B01J2219/00596B01J2219/00722B01J19/0046C12N15/1093
Inventor 安德烈斯·费尔南德斯皮埃尔·F·尹德穆勒尤金·P·马什威廉·巴尼亚伊比尔·J·佩克
Owner TWIST BIOSCI
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