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.

Keywords

BiologyComputer scienceLibrary sciencePhysics

Affiliated Institutions

Related Publications

IN VITRO MUTAGENESIS

TOTAL SYNTHESIS OF MUTANTS . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 424 Solid Phase Chemical Synthe...

1985 Annual Review of Genetics 334 citations

Mutational specificity of depurination.

The mutagenic consequences of damage to DNA produced by low pH and high temperature have been determined in a forward mutational system capable of detecting all classes of mutag...

1984 Proceedings of the National Academy o... 359 citations

Publication Info

Year
1983
Type
article
Volume
11
Issue
12
Pages
4229-4239
Citations
24
Access
Closed

External Links

Social Impact

Social media, news, blog, policy document mentions

Citation Metrics

24
OpenAlex

Cite This

Cinzia Traboni, Riccardo Cortese, Gennaro Ciliberto et al. (1983). A general method to select for M13 clones carrying base pair substitution mutants constructed<i>in vitro</i>. Nucleic Acids Research , 11 (12) , 4229-4239. https://doi.org/10.1093/nar/11.12.4229

Identifiers

DOI
10.1093/nar/11.12.4229