A highly efficient procedure for site-specific mutagenesis of full-length plasmids using Vent DNA polymerase.

Byrappa S, Gavin DK, Gupta KC
Genome Res (1995), Volume 5, Page 404
PubMed entry


Careful titration of Vent polymerase activity allows efficient ...
Careful titration of Vent polymerase activity allows efficient amplification of full-length plasmids (12 kb). The high processivity and fidelity of this enzyme made oligonucleotide-directed site-specific mutagenesis of plasmids a straight-forward process. Using only two primers, a mutagenic and a complementary, single-base mutants of recombinant plasmids were obtained consistently with > 90% efficiency from a single round of PCR. This procedure also made site-specific deletion, insertion, and several bases mutagenesis facile and efficient.




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