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• W1556904239 abstract "This chapter discusses the polymerase chain reaction (PCR)-based point mutagenesis protocol. PCR involves the selective amplification of a segment of DNA template flanked by two synthetic complementary oligodeoxynucleotide primers by repeated cycles of three basic steps: heat denaturation of the template DNA, annealing of the primers to the template to form stable duplexes, and extension of the 3′ ends of the primers by a DNA polymerase. Each new cycle of PCR amplification gives rise to twice the number of copies of the template DNA from the previous cycle as a result of annealing and extension of these primers by DNA polymerase. One of the powerful applications of the PCR is in site-directed mutagenesis protocol to carry out the structural and functional analysis of genes encoding proteins or regulatory elements. Deletions, insertions, and point mutagenesis can be carried out using the PCR. The two-step PCR mutagenesis method is effective in introducing multiple mutations in a defined region preselected by mixed-site mutagenic oligodeoxynucleotides. Vent DNA polymerase can also be used for mutagenesis procedure except that a lower annealing temperature is required." @default.
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• W1556904239 date "1995-01-01" @default.
• W1556904239 modified "2023-10-14" @default.
• W1556904239 title "Polymerase Chain Reaction-Based Point Mutagenesis Protocol" @default.
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• W1556904239 doi "https://doi.org/10.1016/b978-0-12-765561-1.50027-8" @default.
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