An efficient and economic enhancer mix for PCR

Ralser, Markus; Querfurth, Robert; Warnatz, Hans-Jörg; Lehrach, Hans; Yaspo, Marie-Laure; Krobitsch, Sylvia

doi:10.1016/j.bbrc.2006.06.151

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An efficient and economic enhancer mix for PCR

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Ralser, Markus
Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Querfurth, Robert
Dept. of Computational Molecular Biology (Head: Martin Vingron), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Warnatz, Hans-Jörg
Human Chromosome 21 (Marie-Laure Yaspo), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Lehrach, Hans
Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Yaspo, Marie-Laure
Human Chromosome 21 (Marie-Laure Yaspo), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Krobitsch, Sylvia
Neurodegenerative Disorders (Sylvia Krobitsch), Independent Junior Research Groups (OWL), Max Planck Institute for Molecular Genetics, Max Planck Society;

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Citation

Ralser, M., Querfurth, R., Warnatz, H.-J., Lehrach, H., Yaspo, M.-L., & Krobitsch, S. (2006). An efficient and economic enhancer mix for PCR. Biochemical and Biophysical Research Communications (Orlando, FL), 347(3), 747-751. doi:10.1016/j.bbrc.2006.06.151.

Cite as: https://hdl.handle.net/11858/00-001M-0000-0010-83B6-C

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.