Immobilized MutS-Mediated Error Removal of Microchip-Synthesized DNA.
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.
School of Life Science, University of Science and Technology of China, No 443 Huangshan Road, Hefei, Anhui, 230026, People's Republic of China.
Wen
W
Wan
School of Life Science, University of Science and Technology of China, No 443 Huangshan Road, Hefei, Anhui, 230026, People's Republic of China.
Dongmei
D
Wang
School of Life Science, University of Science and Technology of China, No 443 Huangshan Road, Hefei, Anhui, 230026, People's Republic of China.|Hefei National Laboratory for Physical Science at the Microscale, Hefei, 230026, Anhui, People's Republic of China.
Xiaolian
X
Gao
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.|Hefei National Laboratory for Physical Science at the Microscale, Hefei, 230026, Anhui, People's Republic of China. hjiong@ustc.edu.cn.
Jiong
J
Hong
D001483:Base Sequence; D003001:Cloning, Molecular; D055767:Immobilized Proteins / Q000737:chemistry*; D051719:MutS DNA Mismatch-Binding Protein / Q000737:chemistry*; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000737:chemistry
D055767:Immobilized Proteins; D009841:Oligonucleotides; D011993:Recombinant Fusion Proteins; D051719:MutS DNA Mismatch-Binding Protein
De novo gene synthesis; Error removal; Gene assemble; Microchip-synthesized oligonucleotides; MutS