title: 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 degrees 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.
authors.affiliation: Trilink BioTechnologies, Inc., 9955 Mesa Rim Road, San Diego, CA 92121, USA.
authors.forename: Inna
authors.initials: I
authors.lastname: Koukhareva
authors.forename: Huang
authors.initials: H
authors.lastname: Haoqiang
authors.forename: Joyclyn
authors.initials: J
authors.lastname: Yee
authors.forename: Jonathan
authors.initials: J
authors.lastname: Shum
authors.forename: Natasha
authors.initials: N
authors.lastname: Paul
authors.forename: Richard I
authors.initials: RI
authors.lastname: Hogrefe
authors.forename: Alexandre V
authors.initials: AV
authors.lastname: Lebedev
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics; D004279:DNA, Viral / Q000096:biosynthesis; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics; D006358:Hot Temperature; D006801:Humans; D007700:Kinetics; D016133:Polymerase Chain Reaction
chemical_list: D004279:DNA, Viral; D003854:Deoxyribonucleotides; D004259:DNA-Directed DNA Polymerase