PCR-mediated chemical mutagenesis of cloned duplex DNAs

Biotechniques. 1991 Aug;11(2):204-6, 208, 210-1.

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

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Cloning, Molecular
  • DNA / drug effects
  • Humans
  • Molecular Sequence Data
  • Mutagenesis*
  • Nitrous Acid / pharmacology
  • Polymerase Chain Reaction / methods*
  • Ribosomal Proteins / genetics

Substances

  • Ribosomal Proteins
  • ribosomal protein S14
  • DNA
  • Nitrous Acid