Article
PCR-mediated generation of a gene disruption construct without the use of DNA ligase and plasmid vectors.
Nucleic acids research - 15 Jan 2002
Kuwayama Hidekazu, Obara Shinji, Morio Takahiro, Katoh Mariko, Urushihara Hideko, Tanaka Yoshimasa
Abstract excerpt
We introduce a PCR-based procedure for generating a gene disruption construct. This method depends on DNA fragment fusion by the PCR technique and requires only two steps of PCR to obtain a sufficient amount of the gene disruption construct for one transformation experiment. The first step involves three separate PCR syntheses of a selectable marker cassette and the 5'- and 3'-regions of a target gene. Of the...
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