A general method to select for M13 clones carrying base pair substitution mutants constructed in vitro

Cinzia Traboni, Riccardo Cortese, Gennaro Ciliberto, Gianni Cesareni

Research output: Contribution to journalArticle

Abstract

In this paper we describe a method to select base pair substitution mutants constructed in vitro. The mutagenesis is performed by forcing mistakes during in vitro synthesis from a primer annealed to a single stranded DNA template. The selection is based on the fact that, following transformation, the progeny of the strand elongated in vitro and the template strand have different phenotypes. The method is general and applicable to any DNA segment; the type of base pair substitution and its position can be chosen at will. The combined efficiency of mutagenesis and selection allows for 85% frequency of mutants in all analyzed clones.

Original languageEnglish
Pages (from-to)4229-4239
Number of pages11
JournalNucleic Acids Research
Volume11
Issue number12
DOIs
Publication statusPublished - Jun 25 1983

Fingerprint

Mutagenesis
Clone
Mutant
Base Pairing
Substitution
DNA
Substitution reactions
Clone Cells
Template
Single-Stranded DNA
Phenotype
Forcing
Synthesis
In Vitro Techniques

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

A general method to select for M13 clones carrying base pair substitution mutants constructed in vitro. / Traboni, Cinzia; Cortese, Riccardo; Ciliberto, Gennaro; Cesareni, Gianni.

In: Nucleic Acids Research, Vol. 11, No. 12, 25.06.1983, p. 4229-4239.

Research output: Contribution to journalArticle

Traboni, Cinzia ; Cortese, Riccardo ; Ciliberto, Gennaro ; Cesareni, Gianni. / A general method to select for M13 clones carrying base pair substitution mutants constructed in vitro. In: Nucleic Acids Research. 1983 ; Vol. 11, No. 12. pp. 4229-4239.
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