Preparation of modified oligonucleotides by nicking enzyme amplification reaction

0429023 - ÚOCHB 2015 CZ eng C - Conference Paper (international conference)
Ménová, Petra - Hocek, Michal
Preparation of modified oligonucleotides by nicking enzyme amplification reaction.
Chemistry of Nucleic Acid Components. 16th Symposium. Praha: Institute of Organic Chemistry and Biochemistry AS CR, v. v. i, 2014 - (Hocek, M.), s. 324-325. Collection Symposium Series, 14. ISBN 978-80-86241-50-0.
[Symposium on Chemistry of Nucleic Acid Components /16./. Český Krumlov (CZ), 08.06.2014-13.06.2014]
R&D Projects: GA ČR GBP206/12/G151; GA ČR GA203/09/0317; GA AV ČR(CZ) IAA400040901
Institutional support: RVO:61388963
Keywords : dNTPs * DNA * oligonucleotides
Subject RIV: CC - Organic Chemistry

A method for the enzymatic synthesis of short (10-22 nt) base-modified oligonucleotides was developed, based on the nicking enzyme amplification reaction. The methodology, including product isolation and purification, was scaled up to nanomolar amounts. The modified oligonucleotides were successfully used as primers in primer extension and PCR, affording primer-modified oligonucleotides and DNA. Two simple and efficient methods for fluorescent labelling of the PCR products were also developed.
Permanent Link: http://hdl.handle.net/11104/0234438