title: Addressing PCR biases in environmental microbiology studies.
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.
authors.affiliation: Department of Microbiology, Eötvös Loránd University, Budapest, Hungary.
authors.forename: Rita
authors.initials: R
authors.lastname: Sipos
authors.forename: Anna
authors.initials: A
authors.lastname: Székely
authors.forename: Sára
authors.initials: S
authors.lastname: Révész
authors.forename: Károly
authors.initials: K
authors.lastname: Márialigeti
mesh_terms: D017931:DNA Primers / Q000235:genetics; D004783:Environmental Microbiology; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*
chemical_list: D017931:DNA Primers