Abstract
We describe an efficient, PCR-mediated protocol for random chemical mutagenesis of cloned duplex DNAs. The method involves a single molecular cloning step and is compatible with a wide variety of recombinant DNA vectors. To illustrate the procedure, we report the nitrous acid mutagenesis of a human ribosomal protein S14 cDNA fragment.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Base Sequence
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Cloning, Molecular
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DNA / drug effects
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Humans
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Molecular Sequence Data
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Mutagenesis*
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Nitrous Acid / pharmacology
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Polymerase Chain Reaction / methods*
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Ribosomal Proteins / genetics
Substances
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Ribosomal Proteins
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ribosomal protein S14
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DNA
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Nitrous Acid