Ontology highlight
ABSTRACT:
SUBMITTER: Wu W
PROVIDER: S-EPMC1178011 | biostudies-literature | 2005 Jul
REPOSITORIES: biostudies-literature
Wu Wu W Jia Zongchao Z Liu Ping P Xie Zhigang Z Wei Qun Q
Nucleic acids research 20050719 13
We have developed a novel three-primer, one-step PCR-based method for site-directed mutagenesis. This method takes advantage of the fact that template plasmid DNA cannot be efficiently denatured at its reannealing temperature (T(ra)), which is otherwise a troublesome problem in regular PCR. Two flanking primers and one mutagenic primer with different melting temperatures (T(m)) are used together in a single PCR tube continuously without any intervention. A single-stranded mutagenic DNA (smDNA) i ...[more]