M13 phage growth and single-stranded DNA preparation.

Abstract

AbstractM13 bacteriophage has a single-stranded DNA (ssDNA) genome, and has proven an extremely useful vector from which to derive single-stranded templates for sequencing and site-directed mutagenesis. During infection of its host cell, the phage DNA replicates as a double-stranded intermediate from which the ssDNA containing phage particles are produced. Infected cells do not lyse, but instead phage particles are continuously released. Cells infected with M13 phage, however, have a longer replication cycle, which means that as the infection proceeds, the areas of slower-growing cells can be visualized as turbid plaques on the lawn of unaffected E. coli (1). Recombinant M13 phage can be cloned from well-separated plaques and used as a source of SSDNA.KeywordsPhage ParticleIsoamyl AlcoholTurbid PlaquePhage PelletInfection ProceedsThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.