Article
Site-directed mutagenesis of double-stranded DNA by the polymerase chain reaction.
Gene - 30 Dec 1994
Weiner M P, Costa G L, Schoettlin W, Cline J, Mathur E, Bauer J C
Abstract excerpt
We have developed a facile procedure for rapid PCR-based site-directed mutagenesis of double-stranded DNA. Increasing the initial template concentration and decreasing the PCR cycles to 5-10 allows us to reduce the rate of undesired second-site mutations and dramatically increase the time savings. Following PCR, DpnI treatment is used to select against parental DNA molecules. The DpnI (target sequence 5'-Gm6ATC)...
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