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Optimization of DNA shuffling for high fidelity recombination

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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Abstract

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.


Item Type:Article
Additional Information:Copyright © 1997 by Oxford University Press. Received December 19, 1996; Accepted January 27, 1997.
Subject Keywords:in-vitro, fragmentation, polymerases, evolution
Record Number:CaltechAUTHORS:ZHAnar97
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:ZHAnar97
Alternative URL:http://dx.doi.org/10.1093/nar/25.6.1307
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:1215
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:05 Jan 2006
Last Modified:26 Dec 2012 08:43

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