title: Gene synthesis method based on overlap extension PCR and DNAWorks program.
abstract: Gene synthesis by chemical methods provides a powerful tool for modifying genes and exploring their structure, expression, and function in the post-genomic era. However, a bottleneck in recent gene synthesis technologies is the high cost of oligonucleotide synthesis and post-synthesis sequencing. Here, we describe a simple, rapid, and low-cost gene synthesis method based on overlap extension PCR (OE-PCR) and the DNAWorks program. This method enables DNA sequences with sizes ranging from 200 bp to 3 kb to be synthesized with few errors, and these errors can be easily corrected by site-directed mutagenesis. Thus, it is amenable to automation for the multiplexed synthesis of different genes and has a potential for high-throughput gene synthesis.
authors.affiliation: State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Li
authors.forename: Bing-Xue
authors.initials: BX
authors.lastname: Dong
authors.forename: Yu-Huan
authors.initials: YH
authors.lastname: Liu
authors.forename: Chang-Jie
authors.initials: CJ
authors.lastname: Li
authors.forename: Li-Ping
authors.initials: LP
authors.lastname: Zhang
mesh_terms: D019295:Computational Biology / Q000379:methods*; D004247:DNA / Q000138:chemical synthesis*; D005813:Genes, Synthetic; D005818:Genetic Engineering / Q000379:methods; D020407:Internet; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction / Q000379:methods; D012984:Software; D058615:Synthetic Biology / Q000379:methods
chemical_list: D004247:DNA