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Methods Mol Biol. 2017;1472:217-35. doi: 10.1007/978-1-4939-6343-0_17.

Immobilized MutS-Mediated Error Removal of Microchip-Synthesized DNA.

Wan W1, Wang D1, Gao X1,2, Hong J3,4.

Author information

1
School of Life Science, University of Science and Technology of China, No 443 Huangshan Road, Hefei, Anhui, 230026, People's Republic of China.
2
Hefei National Laboratory for Physical Science at the Microscale, Hefei, 230026, Anhui, People's Republic of China.
3
School of Life Science, University of Science and Technology of China, No 443 Huangshan Road, Hefei, Anhui, 230026, People's Republic of China. hjiong@ustc.edu.cn.
4
Hefei National Laboratory for Physical Science at the Microscale, Hefei, 230026, Anhui, People's Republic of China. hjiong@ustc.edu.cn.

Abstract

Applications of microchip-synthesized oligonucleotides for de novo gene synthesis are limited primarily by their high error rates. The mismatch binding protein MutS, which can specifically recognize and bind to mismatches, is one of the cheapest tools for error correction of synthetic DNA. Here, we describe a protocol for removing errors in microchip-synthesized oligonucleotides and for the assembly of DNA segments using these oligonucleotides. This protocol can also be used in traditional de novo gene DNA synthesis.

KEYWORDS:

De novo gene synthesis; Error removal; Gene assemble; Microchip-synthesized oligonucleotides; MutS

PMID:
27671944
DOI:
10.1007/978-1-4939-6343-0_17
[Indexed for MEDLINE]

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