Two-step total gene synthesis method.
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.
Young
Lei
L
Department of Medicine, University of Sydney, Australia. lyoung@student.usyd.edu.au
Dong
Qihan
Q
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
D003062:Codon; D009841:Oligonucleotides; D011994:Recombinant Proteins; D011384:Proinsulin