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Nucleic Acids Research Advance Access originally published online on July 7, 2008
Nucleic Acids Research 2008 36(15):e93; doi:10.1093/nar/gkn421
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Nucleic Acids Research, 2008, Vol. 36, No. 15 e93
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Tetraalkylammonium derivatives as real-time PCR enhancers and stabilizers of the qPCR mixtures containing SYBR Green I

Gouse M. Shaik, Lubica Dráberová, Peter Dráber, Michael Boubelík and Petr Dráber*

Department of Signal Transduction, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Vídenská 1083, 14220 Prague 4, Czech Republic

*To whom correspondence should be addressed. Tel: +420 241062468; Fax: +420 241062214; Email: draberpe{at}img.cas.cz

Received May 1, 2008. Revised June 14, 2008. Accepted June 18, 2008.

Tetraalkylammonium (TAA) derivatives have been reported to serve as stabilizers of asymmetrical cyanine dyes in aqueous solutions and to increase the yield and efficiency of polymerase chain reaction (PCR) detected by end-point analysis. In this study, we compared the ability of various TAA derivatives (with alkyl chain ranging from 1 to 5 carbons) and some other compounds to serve as enhancers of real-time PCR based on fluorescence detection from intercalating dye SYBR Green I (SGI). Our data indicate that TAA chlorides and some other TAA derivatives serve as potent enhancers of SGI-monitored real-time PCR. Optimal results were obtained with 10–16 mM tetrapropylammonium chloride. The effect of TAA compounds was dependent on the nature of counter ions present and composition of the reaction mixtures used. Based on measurements of SGI-generated fluorescence signal in the presence of PCR-amplified DNA fragments, oligonucleotide primers and/or various additives, we propose that TAA-derivatives reduce the binding of SGI to oligonucleotide primers and thus enhance primer–template interactions during annealing phase. Furthermore, these compounds serve as stabilizers of SGI-containing PCR mixtures. The combined data indicate that TAA derivatives might be a new class of additives contributing to robustness of real-time PCR monitored by asymmetrical cyanine dye SGI.


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