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. 1995 Feb;3(1):1-7.
doi: 10.1007/BF02821329.

Site-directed mutagenesis by double polymerase chain reaction

S Barik  1
Affiliations

Site-directed mutagenesis by double polymerase chain reaction

S Barik. Mol Biotechnol. 1995 Feb.

Abstract

A simple and versatile method of in vitro site-specific mutagenesis based on polymerase chain reaction (PCR) is described. The complete method required the use of three oligonucleotide primers and two PCRs. The product of the first PCR was used as one of the primers (megaprimer) in the second PCR. Essentially 100% of the final product incorporated the desired mutation. The various aspects of the procedure and its application is described in detail.

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References

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