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- W2126753602 abstract "We describe a simple protocol for introducing random point mutations into cloned DNA fragments via forced misincorporation of deoxynucleoside triphosphates (dNTPs) by either of two polymerases which lack proofreading activity, reverse transcriptase or a mutant T7 DNA polymerase. A high ratio of one nucleotide to the other three is used to enhance the error rate. Mutagenesis is initiated from a specific primer and restricted to a relatively short segment by limiting the amounts of the nonmutagenic dNTPs. Our method is highly efficient, resulting in the isolation of greater than 50% mutant plasmids with most polymerase/forcing dNTP combinations. A wide spectrum of mutations can be obtained, in contrast to commonly used methods for random mutagenesis." @default.
- W2126753602 created "2016-06-24" @default.
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- W2126753602 date "1990-03-01" @default.
- W2126753602 modified "2023-09-26" @default.
- W2126753602 title "A simple high-efficiency method for random mutagenesis of cloned genes using forced nucleotide misincorporation" @default.
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- W2126753602 doi "https://doi.org/10.1016/0378-1119(90)90066-z" @default.
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