SpringerProtocols: Abstract: Two-Stage Polymerase Chain Reaction Protocol Allowing Introduction of Multiple Mutations, Deletions, and Insertions, Using QuikChangeTM Site-Directed Mutagenesis

Two-Stage Polymerase Chain Reaction Protocol Allowing Introduction of Multiple Mutations, Deletions, and Insertions, Using QuikChangeTM Site-Directed Mutagenesis

By: Wenyan Wang², Bruce A. Malcolm³

Abstract

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The invention of polymerase chain reaction (PCR) (1,2) provided a powerful tool to modify DNA sequences in genetic engineering. With numerous mutagenesis methods available, such as traditional sequential PCR (3), “megaprimer PCR”(4–7), marker-coupled PCR (8), and so on, introducing changes to DNA sequences has become less tedious and more efficient. Recently, marketed site-directed mutagenesis (SDM) kits, such as Transformer™ Site-Directed Mutagenesis Kit (Clontech, San Francisco, CA), and Altered Site® II in vitro Site-Directed Mutagenesis Systems (Promega, Madison, WI), even eliminate the necessity of subcloning the amplified fragment.

Affiliation(s): (2) Department of Structural Chemistry, Schering-Plough Research Institute, Kenilworth, NJ
(3) Department of Antiviral Therapy, Schering-Plough Research Institute, Kenilworth, NJ

Book Title: In Vitro Mutagenesis Protocols

Series: Methods in Molecular Biology | Volume: 182 | Pub. Date: Nov-06-2001 | Page Range: 37-43 | DOI: 10.1385/1-59259-194-9:037

Subject: Genetics/Genomics

References

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