A method for synthesizing texts required for DNA storage based on solid-phase chemical synthesis

A technology of solid-phase chemical synthesis and DNA sequence, which is applied in the field of DNA storage, can solve the problems of cumbersome steps in synthesizing DNA text, increasing DNA text, and low storage capacity, so as to achieve easy portability and follow-up operation, reduce workload, and increase storage capacity Effect

Active Publication Date: 2022-05-03
NANJING ORIGINALCODE PARTNERSHIP (LLP) +1
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0007] The purpose of the present invention is to solve the problems of cumbersome steps for synthesizing DNA text in the process of DNA storage, further purification of the synthesized samples, low storage capacity, and high cost. The present invention provides a synthetic DNA storage method based on solid-phase chemical synthesis. The method of the required text, this method removes the capping operation process in the original solid-phase chemical synthesis, on the one hand simplifies the steps of the traditional phosphite amidotriester method DNA synthesis, on the other hand can greatly increase the synthesis yield, thereby increasing the required DNA text for DNA synthesis

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  • A method for synthesizing texts required for DNA storage based on solid-phase chemical synthesis
  • A method for synthesizing texts required for DNA storage based on solid-phase chemical synthesis
  • A method for synthesizing texts required for DNA storage based on solid-phase chemical synthesis

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Embodiment 1

[0047] In this example, a nucleic acid sequence of 10 bases is used as the information storage sequence, and the DNA sequence required for storage is synthesized by the optimized solid-phase phosphorous triester method.

[0048] (1) In situ synthesis of a nucleic acid sequence based on a solid phase carrier:

[0049] A 10-base nucleic acid sequence (shown in SEQ NO.1) was designed and synthesized, and the synthetic sequence was immobilized on controlled-pore glass beads (CPG) by in situ synthesis.

[0050] The method adopted in this embodiment can operate on batch sequences, therefore, multiple template sequences can be immobilized on the array holes of the chip or other carriers, such as figure 1 shown.

[0051] 5′-AGCT AGCT AG-3′ (SEQ NO.1)

[0052] (2) Refer to Figure 4 As shown, the specific steps of DNA chemical synthesis are as follows:

[0053]

[0054] Wherein, the ratio of each material in the coupling reaction in the fifth step is as follows: Nucleotide on th...

Embodiment 2

[0058] Example 2: Using micron titanium nitride TiN as a carrier for DNA synthesis.

[0059] 1. Activation: Weigh 1g of micron titanium nitride TiN powder in three test tubes, and then shake the mixed solution of water: hydrogen peroxide: concentrated ammonia water = 5:1:1 (ml) at room temperature for 5, 10, and 15 minutes respectively;

[0060] 2. Washing: wash with pure water (pH<8.0, generally wash with a large amount of water 4-5 times, pH can reach 7.5), then wash with ethanol twice, wash with toluene twice, and dry at 80°C for 30 minutes;

[0061] 3. Preparation for the carrier: place the above-mentioned dry micron titanium nitride powder in a mixture of 5ml toluene + 250μl hydroxysilane + 25μl n-butylamine for 4 hours at room temperature, then wash with toluene 5 times and dry in an oven at 110°C 40min to obtain silanized micron titanium nitride powder;

[0062] 4. DNA chain synthesis reaction: operate in a glove box: the above silanized micron titanium nitride powder ...

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Abstract

The invention provides a method for synthesizing text required for DNA storage based on a solid-phase chemical synthesis method, which is improved on the original solid-phase phosphorous acid amido triester method, by removing the Capping (capping) step in the original method , which simplifies the DNA chemical synthesis steps and shortens the time and cost required for the overall synthesis. In addition, no purification steps are required for the synthesized DNA transcript, which further reduces the workload and cost of the entire process. This method increases the synthetic quantity of DNA text on the carrier and greatly improves the storage capacity per unit area. In the reading process, there is a fault-tolerant mechanism that can correct each other among the DNA text strands synthesized by this synthesis method, and the accuracy of reading is improved through high-throughput parallel reading. Finally, in terms of the selection of synthetic vectors, the synthesis method has no special requirements for synthetic vectors, and all general vectors can be used for the synthesis of DNA text.

Description

technical field [0001] The invention belongs to the technical field of DNA storage, and in particular relates to a method for synthesizing text required for DNA storage based on a solid-phase chemical synthesis method. Background technique [0002] With the rapid development of the Internet and artificial intelligence, mankind has entered the era of big data. The amount of data increasing geometrically has exceeded the limit of existing memory storage capacity, and the storage capacity of traditional storage devices such as magnetic storage (tape, hard drive), optical storage (Blu-ray) and solid-state storage (flash memory) has been unable to meet Explosive storage requirements. Moreover, these traditional storage media have defects in varying degrees in terms of storage density, storage time, and maintenance cost. Therefore, storage media that are different from traditional ones urgently need to be developed. [0003] Nucleic acid (nucleic acid) is the main carrier of th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07H21/04C07H1/00
CPCC07H21/04C07H1/00
Inventor 刘全俊闫汉张振朱立博
Owner NANJING ORIGINALCODE PARTNERSHIP (LLP)
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