A PCR-based strategy for extensive mutagenesis of a target DNA sequence

H. G. Morrison, R. C. Desrosiers

Research output: Contribution to journalArticle

37 Scopus citations

Abstract

A mixed population of mutagenic oligonucleotide primers was used to generate a set of point mutations in a short region of a retroviral gene by PCR amplification. The mixed population of mutagenic primers was generated by incorporating a mixture of A, G, C and T at specific sites during oligonucleotide synthesis. With the proportions of mutagenic nucleotides used for our experiments, 47 percent of the 213 clones analyzed had one or more point mutation in the target DNA sequence. In addition, unpredicted mutations were observed that contributed to the mutagenic complexity of the population. We have found this approach to be an efficient means for extensive mutagenesis of a defined target DNA sequence.

Original languageEnglish (US)
Pages (from-to)454-457
Number of pages4
JournalBioTechniques
Volume14
Issue number3
StatePublished - Jan 1 1993
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

Fingerprint Dive into the research topics of 'A PCR-based strategy for extensive mutagenesis of a target DNA sequence'. Together they form a unique fingerprint.

  • Cite this