Nucleic Acids Research, Vol 25, Issue 6 1307-1308, Copyright © 1997 by Oxford University Press
H Zhao and FH Arnold
A convenient 'DNA shuffling' protocol for random recombination of
homologous genes in vitro with a very low rate of associated point
mutagenesis (0.05%) is described. In addition, the mutagenesis rate can be
controlled over a wide range by the inclusion of Mn2+or Mg2+during DNase I
digestion, by choice of DNA polymerase used during gene reassembly as well
as how the genes are prepared for shuffling (PCR amplification versus
restriction enzyme digestion of plasmid DNA). These protocols should be
useful for in vitro protein evolution, for DNA based computing and for
structure-function studies of evolutionarily related genes.
ARTICLES
Optimization of DNA shuffling for high fidelity recombination
Division of Chemistry and Chemical Engineering 210-41, California Institute of Technology, Pasadena, CA 91125, USA.
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