An improved method of gene synthesis based on DNA works software and overlap extension PCR.

An improved method of gene synthesis based on DNA works software and overlap extension PCR. Research Abstract Details 

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An improved method of gene synthesis based on DNA works software and overlap extension PCR. Abstract Text:

Bingxue Dong,Runqian Mao,Baojian Li,Qiuyun Liu,Peilin Xu,Gang Li,Bingxue Dong,Runqian Mao,Baojian Li,Qiuyun Liu,Peilin Xu,Gang Li,

A bottleneck in recent gene synthesis technologies is the high cost of oligonucleotide synthesis and post-synthesis sequencing. In this article, a simple and rapid method for low-cost gene synthesis technology was developed based on DNAWorks program and an improved single-step overlap extension PCR (OE-PCR). This method enables any DNA sequence to be synthesized with few errors, then any mutated sites could be corrected by site-specific mutagenesis technology or PCR amplification-assembly method, which can amplify different DNA fragments of target gene followed by assembly into an entire gene through their overlapped region. Eventually, full-length DNA sequence without error was obtained via this novel method. Our method is simple, rapid and low-cost, and also easily amenable to automation based on a DNAWorks design program and defined set of OE-PCR reaction conditions suitable for different genes. Using this method, several genes including Manganese peroxidase gene (Mnp) of Phanerochaete chrysosporium (P. chrysosporium), Laccase gene (Lac) of Trametes versicolor (T. versicolor) and Cip1 peroxidase gene (cip 1) of Coprinus cinereus (C. cinereus) with sizes ranging from 1.0 kb to 1.5 kb have been synthesized successfully.

An improved method of gene synthesis based on DNA works software and overlap extension PCR. Publishing Authors By Initials

B Dong,R Mao,B Li,Q Liu,P Xu,G Li,B Dong,R Mao,B Li,Q Liu,P Xu,G Li,

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An improved method of gene synthesis based on DNA works software and overlap extension PCR. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Molecular biotechnology

VOLUME: 37

Page Numbers: 195-200

Journal Abbreviation: Mol. Biotechnol.

ISSN: 1073-6085

DAY: 3

MONTH: 08

YEAR: 2007

An improved method of gene synthesis based on DNA works software and overlap extension PCR. Information

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LANGUAGE: eng

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Grant and Affiliation Information for An improved method of gene synthesis based on DNA works software and overlap extension PCR.

AFFILIATION: The Key Laboratory of Gene Engineering of Ministry of Education, Sun Yat-sen University, Guangzhou, P.R. China.

Country: United States

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MEDLINETA: Mol Biotechnol

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