1,2-propanediol-trehalose mixture as a potent quantitative real-time PCR enhancer

0365065 - ÚMG 2012 RIV GB eng J - Journal Article
Horáková, Helena - Polakovičová, Iva - Shaik, Gouse Mohiddin - Eitler, Jiří - Bugajev, Viktor - Dráberová, Lubica - Dráber, Petr
1,2-propanediol-trehalose mixture as a potent quantitative real-time PCR enhancer.
BMC Biotechnology. Roč. 11, - (2011), e41. E-ISSN 1472-6750
R&D Projects: GA AV ČR KAN200520701; GA ČR(CZ) GD204/09/H084; GA ČR GA301/09/1826; GA ČR GAP302/10/1759
Grant - others:GA AV CR(CZ) M200520901
Institutional research plan: CEZ:AV0Z50520514
Keywords : qPCR enhancer * trehalose * 1,2-propanediol
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 2.349, year: 2011

Quantitative real-time PCR (qPCR) is becoming increasingly important for DNA genotyping and gene expression analysis. Buffers of different composition are used for various DNA dyes used in qPCR. In search for a PCR mix compatible with variety of DNA dyes, and suitable for efficient amplification of difficult-to-amplify DNA templates, such as those in whole blood, of medium size and/or GC-rich, we found excellent performance of a PCR mix supplemented with 1 M 1,2-propanediol and 0.2 M trehalose (PT enhancer). These two additives together decreased DNA melting temperature and efficiently neutralized PCR inhibitors present in blood samples. They also made possible more efficient amplification of GC-rich templates than betaine and other previously described additives. The data indicate that PCR mixes supplemented with PT enhancer are suitable for DNA amplification in the presence of various DNA dyes and for a variety of templates which otherwise can be amplified with difficulty.
Permanent Link: http://hdl.handle.net/11104/0200401