Convenient modification of the method for oligonucleotide-directed in vitro mutagenesis of cloned DNA.
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%.
D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics* / Q000378:metabolism; D004274:DNA, Recombinant; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005090:Exodeoxyribonucleases / Q000378:metabolism; D009154:Mutation; D009692:Nucleic Acid Heteroduplexes; D009838:Oligodeoxyribonucleotides / Q000235:genetics*; D009841:Oligonucleotides / Q000235:genetics*; D010957:Plasmids; D014169:Transformation, Bacterial
D004269:DNA, Bacterial; D004274:DNA, Recombinant; D009692:Nucleic Acid Heteroduplexes; D009838:Oligodeoxyribonucleotides; D009841:Oligonucleotides; D004259:DNA-Directed DNA Polymerase; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III