title: Two-step total gene synthesis method.
abstract: In the post-genomic era, the ability to synthesize any arbitrary DNA sequence is increasingly in demand. A bottleneck in current gene synthesis technologies is the associated cost, due primarily to the high cost of oligonucleotides synthesis and post-synthesis sequencing. In the present paper, an improved method for low-cost gene synthesis that combines dual asymmetrical PCR and overlap extension PCR is presented, which enables any DNA sequence to be synthesized error free. Additionally, the method is easily amenable to automation.
authors.affiliation: Department of Medicine, University of Sydney, Australia. lyoung@student.usyd.edu.au
authors.forename: Lei
authors.initials: L
authors.lastname: Young
authors.forename: Qihan
authors.initials: Q
authors.lastname: Dong
mesh_terms: D003062:Codon / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics; D005813:Genes, Synthetic / Q000235:genetics*; D005821:Genetic Techniques; D009841:Oligonucleotides / Q000138:chemical synthesis / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D011384:Proinsulin / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D015203:Reproducibility of Results
chemical_list: D003062:Codon; D009841:Oligonucleotides; D011994:Recombinant Proteins; D011384:Proinsulin