Synthesis of 3'-O-fluorescently mono-modified reversible terminators and their uses in sequencing-by-synthesis
- Synthesis of 3'-O-fluorescently mono-modified reversible terminators and their uses in sequencing-by-synthesis
- 김다래; 김택수; 김은선; 민선준; 신동윤; 안대로
- Sequencing-by-synthesis; Reversible terminator; Modified nucleotide; DNA polymerase; Triphosphate
- Issue Date
- Bioorganic & medicinal chemistry letters
- VOL 24, NO 1, 209-213
- 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.
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