title: Barcoded primers used in multiplex amplicon pyrosequencing bias amplification.
abstract: "Barcode-tagged" PCR primers used for multiplex amplicon sequencing generate a thus-far-overlooked amplification bias that produces variable terminal restriction fragment length polymorphism (T-RFLP) and pyrosequencing data from the same environmental DNA template. We propose a simple two-step PCR approach that increases reproducibility and consistently recovers higher genetic diversity in pyrosequencing libraries.
authors: Department of Microbial Ecology, Vienna Ecology Center, Faculty of Life Sciences, University of Vienna, Althanstrasse 14, A-1090 Vienna, Austria.
authors: David
authors: D
authors: Berry
authors: Karim
authors: K
authors: Ben Mahfoudh
authors: Michael
authors: M
authors: Wagner
authors: Alexander
authors: A
authors: Loy
mesh_terms: D017931:DNA Primers / Q000235:genetics*; D003951:Diagnostic Errors; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D017931:DNA Primers