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.
Key wordsMutS Microchip-synthesized oligonucleotides Gene assemble Error removal De novo gene synthesis
This work was supported by a grant-in-aid from the National Natural Science Foundation of China (31270149), the National High Technology Research and Development Program (2012AA02A708), Anhui Provincial Natural Science Foundation (1608085MC47),the Fundamental Research Funds for the Central Universities (WK2070000059), the China Postdoctoral Science Foundation (2015M580546). This work also earned technical support from the Core Facility Center for Life Sciences, University of Science and Technology of China.