Addressing PCR biases in environmental microbiology studies.

Rita Sipos, Anna Székely, Sára Révész, Károly Márialigeti

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Each step of a molecular environmental microbiology study is prone to errors, though the qualitative and quantitative biases of PCR amplification could result in the most serious biases. One has to be aware of this fact, and well-characterized PCR biases have to be avoided by using target-optimized PCR protocols. The most important tasks are primer and thermal profile optimization. We have shown that primer mismatches, even in the case of universal primers, can cause almost complete missing of common taxa from clone libraries, for example. Similarly high annealing temperatures can drastically distort community composition of the sample in the PCR product. Strategies of primer selection and PCR thermal profile design are discussed in detail.

Original languageEnglish
Pages (from-to)37-58
Number of pages22
JournalMethods in molecular biology (Clifton, N.J.)
Volume599
DOIs
Publication statusPublished - 2010

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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