Protocol

Synthetic DNA

Volume 1472 of the series Methods in Molecular Biology pp 63-78

Date:

SpeedyGenes: Exploiting an Improved Gene Synthesis Method for the Efficient Production of Synthetic Protein Libraries for Directed Evolution

  • Andrew CurrinAffiliated withManchester Institute of Biotechnology, The University of ManchesterSchool of Chemistry, The University of ManchesterCentre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), The University of Manchester Email author 
  • , Neil SwainstonAffiliated withManchester Institute of Biotechnology, The University of ManchesterCentre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), The University of ManchesterSchool of Computer Science, The University of Manchester
  • , Philip J. DayAffiliated withManchester Institute of Biotechnology, The University of ManchesterCentre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), The University of ManchesterFaculty of Medical and Human Sciences, The University of Manchester
  • , Douglas B. KellAffiliated withManchester Institute of Biotechnology, The University of ManchesterSchool of Chemistry, The University of ManchesterCentre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), The University of Manchester

* Final gross prices may vary according to local VAT.

Get Access

Abstract

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.

Key words

Directed evolution Error correction Gene synthesis Protein libraries Synthetic biology