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

Research output: Contribution to journalArticle

37 Citations (Scopus)

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 - 1993
Externally publishedYes

Fingerprint

Mutagenesis
DNA sequences
Point Mutation
Polymerase Chain Reaction
DNA Primers
Oligonucleotides
Population
Amplification
Nucleotides
Gene Amplification
Genes
Clone Cells
Mutation
Experiments

ASJC Scopus subject areas

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

Cite this

A PCR-based strategy for extensive mutagenesis of a target DNA sequence. / Morrison, H. G.; Desrosiers, Ronald Charles.

In: BioTechniques, Vol. 14, No. 3, 1993, p. 454-457.

Research output: Contribution to journalArticle

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