Wan Wen, Wang Dongmei, Gao Xiaolian, Hong Jiong
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.
Methods Mol Biol. 2017;1472:217-35. doi: 10.1007/978-1-4939-6343-0_17.
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.
微芯片合成寡核苷酸在从头基因合成中的应用主要受到其高错误率的限制。错配结合蛋白MutS能够特异性识别并结合错配，是用于合成DNA纠错的最经济工具之一。在此，我们描述了一种用于去除微芯片合成寡核苷酸中的错误以及使用这些寡核苷酸组装DNA片段的方案。该方案也可用于传统的从头基因DNA合成。
