PCR-mediated chemical mutagenesis of cloned duplex DNAs.
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.
Diaz
J J
JJ
Division of Biology, Kansas State University, Manhattan 66506.
Rhoads
D D
DD
Roufa
D J
DJ
D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009608:Nitrous Acid / Q000494:pharmacology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012269:Ribosomal Proteins / Q000235:genetics
D012269:Ribosomal Proteins; C039747:ribosomal protein S14; D004247:DNA; D009608:Nitrous Acid