Protocol

Synthetic DNA

Volume 1472 of the series Methods in Molecular Biology pp 217-235

Date:

Immobilized MutS-Mediated Error Removal of Microchip-Synthesized DNA

  • Wen WanAffiliated withSchool of Life Science, University of Science and Technology of China
  • , Dongmei WangAffiliated withSchool of Life Science, University of Science and Technology of China
  • , Xiaolian GaoAffiliated withSchool of Life Science, University of Science and Technology of ChinaHefei National Laboratory for Physical Science at the Microscale
  • , Jiong HongAffiliated withSchool of Life Science, University of Science and Technology of ChinaHefei National Laboratory for Physical Science at the Microscale Email author 

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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.

Key words

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