SpeedyGenes: Exploiting an Improved Gene Synthesis Method for the Efficient Production of Synthetic Protein Libraries for Directed Evolution.
Gene synthesis is a fundamental technology underpinning much research in the life sciences. In particular, synthetic biology and biotechnology utilize gene synthesis to assemble any desired DNA sequence, which can then be incorporated into novel parts and pathways. Here, we describe SpeedyGenes, a gene synthesis method that can assemble DNA sequences with greater fidelity (fewer errors) than existing methods, but that can also be used to encode extensive, statistically designed sequence variation at any position in the sequence to create diverse (but accurate) variant libraries. We summarize the integrated use of GeneGenie to design DNA and oligonucleotide sequences, followed by the procedure for assembling these accurately and efficiently using SpeedyGenes.
Manchester Institute of Biotechnology, The University of Manchester, 131, Princess St, Manchester, M1 7DN, UK. andrew.currin@manchester.ac.uk.|School of Chemistry, The University of Manchester, Manchester, M13 9PL, UK. andrew.currin@manchester.ac.uk.|Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), The University of Manchester, 131, Princess St, Manchester, M1 7DN, UK. andrew.currin@manchester.ac.uk.
Andrew
A
Currin
Manchester Institute of Biotechnology, The University of Manchester, 131, Princess St, Manchester, M1 7DN, UK.|Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), The University of Manchester, 131, Princess St, Manchester, M1 7DN, UK.|School of Computer Science, The University of Manchester, Manchester, M13 9PL, UK.
Neil
N
Swainston
Manchester Institute of Biotechnology, The University of Manchester, 131, Princess St, Manchester, M1 7DN, UK.|Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), The University of Manchester, 131, Princess St, Manchester, M1 7DN, UK.|Faculty of Medical and Human Sciences, The University of Manchester, Manchester, M13 9PT, UK.
Philip J
PJ
Day
Manchester Institute of Biotechnology, The University of Manchester, 131, Princess St, Manchester, M1 7DN, UK.|School of Chemistry, The University of Manchester, Manchester, M13 9PL, UK.|Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), The University of Manchester, 131, Princess St, Manchester, M1 7DN, UK.
Douglas B
DB
Kell
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D019020:Directed Molecular Evolution; D004926:Escherichia coli / Q000235:genetics; D005813:Genes, Synthetic; D009841:Oligonucleotides / Q000235:genetics; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D014169:Transformation, Bacterial; D014584:User-Computer Interface
D003062:Codon; D009841:Oligonucleotides; D019151:Peptide Library
Directed evolution; Error correction; Gene synthesis; Protein libraries; Synthetic biology