Zhao, Huimin and Arnold, Frances H. (1997) Optimization of DNA shuffling for high fidelity recombination. Nucleic Acids Research, 25 (6). pp. 1307-1308. ISSN 0305-1048 http://resolver.caltech.edu/CaltechAUTHORS:ZHAnar97
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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.
|Additional Information:||Copyright © 1997 by Oxford University Press. Received December 19, 1996; Accepted January 27, 1997.|
|Subject Keywords:||in-vitro, fragmentation, polymerases, evolution|
|Usage Policy:||No commercial reproduction, distribution, display or performance rights in this work are provided.|
|Deposited By:||Tony Diaz|
|Deposited On:||05 Jan 2006|
|Last Modified:||26 Dec 2012 08:43|
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