Optimization of DNA shuffling for high fidelity recombination

Huimin Zhao, Frances H. Arnold

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)1307-1308
Number of pages2
JournalNucleic acids research
Volume25
Issue number6
DOIs
StatePublished - Mar 15 1997
Externally publishedYes

Fingerprint

DNA Shuffling
Genetic Recombination
Mutagenesis
Genes
Digestion
Deoxyribonuclease I
DNA
DNA-Directed DNA Polymerase
Plasmids
Polymerase Chain Reaction
Enzymes
Proteins
In Vitro Techniques

ASJC Scopus subject areas

  • Genetics

Cite this

Optimization of DNA shuffling for high fidelity recombination. / Zhao, Huimin; Arnold, Frances H.

In: Nucleic acids research, Vol. 25, No. 6, 15.03.1997, p. 1307-1308.

Research output: Contribution to journalArticle

Zhao, Huimin ; Arnold, Frances H. / Optimization of DNA shuffling for high fidelity recombination. In: Nucleic acids research. 1997 ; Vol. 25, No. 6. pp. 1307-1308.
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