title: An efficient and economic enhancer mix for PCR.
abstract: Polymerase chain reaction (PCR) has become a fundamental technique in molecular biology. Nonetheless, further improvements of the existing protocols are required to broaden the applicability of PCR for routine diagnostic purposes, to enhance the specificity and the yield of PCRs as well as to reduce the costs for high-throughput applications. One known problem typically reported in PCR experiments is the poor amplification of GC-rich DNA sequences. Here we designed and tested a novel effective and low-cost PCR enhancer, a concentration-dependent combination of betaine, dithiothreitol, and dimethyl sulfoxide that broadly enhanced the quantitative and/or qualitative output of PCRs. Additionally, we showed that the performances of this enhancer mix are comparable to those of commercially available PCR additives and highly effective with different DNA polymerases. Thus, we propose the routine application of this PCR enhancer mix for low- and high-throughput experiments.
authors.affiliation: Max Planck Institute for Molecular Genetics, Berlin, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Ralser
authors.forename: Robert
authors.initials: R
authors.lastname: Querfurth
authors.forename: Hans-Jörg
authors.initials: HJ
authors.lastname: Warnatz
authors.forename: Hans
authors.initials: H
authors.lastname: Lehrach
authors.forename: Marie-Laure
authors.initials: ML
authors.lastname: Yaspo
authors.forename: Sylvia
authors.initials: S
authors.lastname: Krobitsch
mesh_terms: D004742:Enhancer Elements, Genetic / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000191:economics* / Q000379:methods*; D012996:Solutions
chemical_list: D012996:Solutions