Optimization of DNA shuffling for high fidelity recombination | Zendy

Huimin Zhao | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

Optimization of DNA shuffling for high fidelity recombination

Author(s) -

Huimin Zhao

Publication year - 1997

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/25.6.1307

Subject(s) - dna shuffling , biology , mutagenesis , in vitro recombination , genetics , dna , gene , shuffling , plasmid , homologous recombination , microbiology and biotechnology , restriction enzyme , mutation , directed evolution , molecular cloning , mutant , peptide sequence , computer science , programming language

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.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research