Synthesis of 3 '-O-fluorescently mono-modified reversible terminators and their uses in sequencing-by-synthesis

Authors
Kim, Da-RaeKim, Taek-SooKim, EunsunMin, Sun-JoonShin, DongyunAhn, Dae-Ro
Issue Date
2014-01-01
Publisher
Pergamon Press Ltd.
Citation
Bioorganic & Medicinal Chemistry Letters, v.24, no.1, pp.209 - 213
Abstract
Next-generation sequencing (NGS) technologies recently developed are now used for study of genomes from various organisms. Sequencing-by-synthesis (SBS) is a key strategy in the NGS. The SBS uses nucleotides so-called dual-modified reversible terminators (DRTs) in which bases are labeled with fluorophores and 3'-OH is protected with a reversibly cleavable chemical group, respectively. In this study, we examined the possibility of performing SBS with mono-modified reversible terminators (MRTs), in which the reversible blocking group on the 3'-OH plays a dual role as a fluorescent signal report as well as a chemical protection. We studied cyclic reversible termination by using two MRTs (dA and dT), wherein the modifications were two different fluorophores and cleavable to regenerate a free 3'-OH. We here demonstrated that SBS could be achieved with incorporation of MRTs by a DNA polymerase and correct base-calls based on the two different colors from the fluorophores. (C) 2013 Elsevier Ltd. All rights reserved.
Keywords
DNA-POLYMERASE; TRIPHOSPHATES; NUCLEOTIDES; DNA-POLYMERASE; TRIPHOSPHATES; NUCLEOTIDES; Sequencing-by-synthesis; Reversible terminator; Modified nucleotide; DNA polymerase; Triphosphate
ISSN
0960-894X
URI
https://pubs.kist.re.kr/handle/201004/127239
DOI
10.1016/j.bmcl.2013.11.040
Appears in Collections:
KIST Article > 2014
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