Revealing biases inherent in recombination protocols |
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| Authors: | Javier F Chaparro-Riggers Bernard LW Loo Karen M Polizzi Phillip R Gibbs Xiao-Song Tang Mark J Nelson Andreas S Bommarius |
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| Affiliation: | (1) School of Chemical and Biomolecular Engineering, Parker H. Petit Institute of Bioengineering and Bioscience, School of Chemistry and Biochemistry, Georgia Institute of Technology, 315 Ferst Drive, Atlanta, GA 30332-0363, USA;(2) School of Chemistry and Biochemistry, Georgia Institute of Technology, 315 Ferst Drive, Atlanta, GA 30332-0363, USA;(3) EI DuPont de Nemours & Company, PO Box 80328, Wilmington, DE 19880-0328, USA;(4) Stheno Corporation, 311 Ferst Drive, Atlanta, 30332-0100, USA |
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| Abstract: | Background The recombination of homologous genes is an effective protein engineering tool to evolve proteins. DNA shuffling by gene fragmentation and reassembly has dominated the literature since its first publication, but this fragmentation-based method is labor intensive. Recently, a fragmentation-free PCR based protocol has been published, termed recombination-dependent PCR, which is easy to perform. However, a detailed comparison of both methods is still missing. |
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