Real-time quantitative PCR to discriminate and quantify lambdoid bacteriophages of Escherichia coli K-12

Quantification of bacteriophages by real-time quantitative PCR (qPCR) is an interesting alternative to the traditional plaque assay. Importantly, the method should in principle be able to discriminate between closely related phages that are indistinguishable by most other means. Here, a method is pr...

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Bibliographic Details
Published inBacteriophage Vol. 2; no. 2; pp. 98 - 104
Main Author Refardt, Dominik
Format Journal Article
LanguageEnglish
Published United States Taylor & Francis 01.04.2012
Landes Bioscience
Subjects
bacteriophage lambda
bacteriophages
Binding
Biology
Bioscience
Calcium
Cancer
Cell
Cycle
detection
discrimination
Escherichia coli
lambdoid bacteriophages
Landes
Methods and Protocols
multiple infections
Organogenesis
polylysogeny
Proteins
quantification
real-time quantitative PCR
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Summary:Quantification of bacteriophages by real-time quantitative PCR (qPCR) is an interesting alternative to the traditional plaque assay. Importantly, the method should in principle be able to discriminate between closely related phages that are indistinguishable by most other means. Here, a method is presented that employs qPCR to discriminate and quantify ten closely related lambdoid phages of Escherichia coli str. K-12. It is shown that (1) treatment of samples with DNase efficiently removes non-encapsidated DNA, while the titer of plaque forming units is not affected, (2) individual phage types can be accurately quantified in mixed lysates, and (3) the detection limit corresponds to that of a plaque assay. The method is used to quantify individual phage types that are released from lysogens that carry up to three different prophages.
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ISSN:2159-7073
2159-7081
2159-7081
DOI:10.4161/bact.20092