Method for efficiently synthesizing long-fragment and complex-fragment DNA in vitro

An in vitro synthesis and long fragment technology, which is applied in DNA preparation, recombinant DNA technology, biochemical equipment and methods, etc., can solve the problems of synthesis failure, energy and time consumption, short synthesis length, etc., and achieve the effect of stable synthesis quality

Inactive Publication Date: 2016-12-14
WUHAN BIORUN BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Another solution uses chip technology for high-throughput DNA synthesis. Although DNA can be synthesized on a large scale and at low cost, the synthesis length is relatively short.
For the synthesis of long-chain complex DNA, and the synthesis of long-chain DNA is still a difficult problem in this field, it takes a lot of energy and time, the synthesis cycle is not guaranteed, and even the synthesis fails

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0019] Example: a 10221bp DNA synthesis

[0020] The first step is to split the entire fragment into two-level fragments

[0021] According to the length of the total sequence, the DNA fragment will be split into 9 segments, each of which is about 1136bp.

[0022] SegB1:

[0023] 1-tagaatagca tcggtaacat gagcaaagtc tgccgcctta caacggctct cccgctgacg-60

[0024] 61-ccgtcccgga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg-120

[0025] 121-gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacaaattgacgcttaga-180

[0026] 181-caacttaataacacattgcggacgtttttaatgttagactgaattaacgccgaattaatt-240

[0027] 241-cgggggatct ggattttagt actggatttt ggttttagga attagaaattttatgatag-300

[0028] 301-aagtatttta caaatacaaa tacataactaa gggtttctta tatgctcaacacatgagcga-360

[0029] 361-aaccctatag gaaccctaat tcccttatct gggaactact caacacattattatggagaaa-420

[0030] 421-ctcgagcttg tcgatcgaca gatccggtcg gcatctactc tatttctttgccctcggacg-480

[0031] 481-agtgctgggg cgtcggtttc cactatcggc gagtacttct aca...

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PUM

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Abstract

The invention provides a method for efficiently synthesizing long-fragment and complex-fragment DNA in vitro. Firstly, primary fragments are synthesized by a primer; secondly, fragments are assembled; finally, the fragments are assembled to the DNA sequence. The DNA fragments synthesized by the method has stable synthetic quality and are not interfered by DNA complexity and GC content; superlong DNA fragments can be synthesized; the DNA fragments with length being within 30 kb can be stably synthesized; the requirements of the modern molecular experiments are met.

Description

technical field [0001] The invention relates to a high-efficiency in vitro synthesis method of long-segment and complex-segment DNA. Background technique [0002] In vitro artificial synthesis of DNA is an experiment often encountered in current biological research and applications. It is generally entrusted to a specialized biological company to synthesize. At present, the most widely used synthesis method is to design multiple overlapping primers through annealing and amplification, and then perform PCR. Amplify to obtain full-length DNA. Another solution uses chip technology for high-throughput DNA synthesis. Although DNA can be synthesized on a large scale and at low cost, the synthesis length is relatively short. The synthesis of long-chain complex DNA, and the synthesis of long-chain DNA is still a difficult problem in this field. It takes a lot of energy and time, and the synthesis cycle is not guaranteed, and even the synthesis fails. Contents of the invention ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/10C12Q2521/301C12Q2521/501
Inventor 李阳
Owner WUHAN BIORUN BIO TECH
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