Heat Activatable 3′-modified dNTPs: Synthesis and Application for Hot Start PCR

Abstract Several 3′-ether and 3′-ester derivatives of 2′-deoxyribonucleoside 5′-triphosphates (dNTPs) were prepared. These dNTP derivatives were not substrates for DNA polymerase and did not support primer extension at room temperature. However, by short pre-heating to 95°C in PCR buffer, these 3′-m...

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Bibliographic Details
Published in:Nucleic Acids Symposium Series 2008, Vol.52 (1), p.259-260
Main Authors: Koukhareva, Inna, Haoqiang, Huang, Yee, Joyclyn, Shum, Jonathan, Paul, Natasha, Hogrefe, Richard I., Lebedev, Alexandre V.
Format: Article
Language:English
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Summary:Abstract Several 3′-ether and 3′-ester derivatives of 2′-deoxyribonucleoside 5′-triphosphates (dNTPs) were prepared. These dNTP derivatives were not substrates for DNA polymerase and did not support primer extension at room temperature. However, by short pre-heating to 95°C in PCR buffer, these 3′-modified dNTPs can be converted to corresponding unmodified natural dNTPs that efficiently support PCR amplification. The analysis of PCR products obtained with 3′-modified dNTPs revealed a significant improvement in PCR performance resulting in higher amplicon yield and reduced formation of off-target products (mis-priming and primer dimer). Among the studied 3′-modified dNTPs, the 3′-tetrahydrofuranyl derivatives showed the best results.
ISSN:0261-3166
1746-8272
DOI:10.1093/nass/nrn131