Convenient modification of the method for oligonucleotide-directed in vitro mutagenesis of cloned DNA

V A Efimov; O V Mirskikh; O G Chakhmakhcheva; Ovchinnikov YuA

doi:10.1016/0014-5793(85)80302-1

Convenient modification of the method for oligonucleotide-directed in vitro mutagenesis of cloned DNA

FEBS Lett. 1985 Feb 25;181(2):407-11. doi: 10.1016/0014-5793(85)80302-1.

Authors

V A Efimov, O V Mirskikh, O G Chakhmakhcheva, Ovchinnikov YuA

PMID: 3882458
DOI: 10.1016/0014-5793(85)80302-1

Abstract

A new modification of the oligonucleotide-mediated mutagenesis technique has been developed. The proposed methodology has been used to produce specific base changes in the double-stranded plasmid DNA. For this purpose, special cloning vectors have been constructed using the synthetic oligodeoxyribonucleotides. The developed method allows the production of mutant DNA from those of the wild-type with a yield of 10-20%.

MeSH terms

Base Sequence
DNA, Bacterial / genetics*
DNA, Bacterial / metabolism
DNA, Recombinant*
DNA-Directed DNA Polymerase / metabolism
Escherichia coli / genetics
Exodeoxyribonucleases / metabolism
Mutation
Nucleic Acid Heteroduplexes
Oligodeoxyribonucleotides / genetics*
Oligonucleotides / genetics*
Plasmids
Transformation, Bacterial

Substances

DNA, Bacterial
DNA, Recombinant
Nucleic Acid Heteroduplexes
Oligodeoxyribonucleotides
Oligonucleotides
DNA-Directed DNA Polymerase
Exodeoxyribonucleases
exodeoxyribonuclease III