Format

Send to

Choose Destination
Methods Mol Biol. 2017;1472:63-78. doi: 10.1007/978-1-4939-6343-0_5.

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

Currin A1,2,3, Swainston N4,5,6, Day PJ4,5,7, Kell DB4,8,5.

Author information

1
Manchester Institute of Biotechnology, The University of Manchester, 131, Princess St, Manchester, M1 7DN, UK. andrew.currin@manchester.ac.uk.
2
School of Chemistry, The University of Manchester, Manchester, M13 9PL, UK. andrew.currin@manchester.ac.uk.
3
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.
4
Manchester Institute of Biotechnology, The University of Manchester, 131, Princess St, Manchester, M1 7DN, UK.
5
Centre for Synthetic Biology of Fine and Speciality Chemicals (SYNBIOCHEM), The University of Manchester, 131, Princess St, Manchester, M1 7DN, UK.
6
School of Computer Science, The University of Manchester, Manchester, M13 9PL, UK.
7
Faculty of Medical and Human Sciences, The University of Manchester, Manchester, M13 9PT, UK.
8
School of Chemistry, The University of Manchester, Manchester, M13 9PL, UK.

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.

KEYWORDS:

Directed evolution; Error correction; Gene synthesis; Protein libraries; Synthetic biology

PMID:
27671932
DOI:
10.1007/978-1-4939-6343-0_5
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center