title: Synthesis of 3'-O-fluorescently mono-modified reversible terminators and their uses in sequencing-by-synthesis.
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.
authors: Center for Theragnosis, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea.
authors: Da-Rae
authors: DR
authors: Kim
authors: College of Pharmacy, Gachon University, 191 Hambakmoero, Yoensu-Gu, Incheon, Republic of Korea.
authors: Taek-Soo
authors: TS
authors: Kim
authors: Center for Neuro-medicine, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea.
authors: Eunsun
authors: E
authors: Kim
authors: Center for Neuro-medicine, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea; Department of Biological Chemistry, University of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea.
authors: Sun-Joon
authors: SJ
authors: Min
authors: College of Pharmacy, Gachon University, 191 Hambakmoero, Yoensu-Gu, Incheon, Republic of Korea. Electronic address: dyshin@gachon.ac.kr.
authors: Dongyun
authors: D
authors: Shin
authors: Center for Theragnosis, Korea Institute of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea; Department of Biological Chemistry, University of Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul 136-791, Republic of Korea. Electronic address: drahn@kist.re.kr.
authors: Dae-Ro
authors: DR
authors: Ahn
mesh_terms: D015394:Molecular Structure; D009711:Nucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D017422:Sequence Analysis, DNA
chemical_list: D009711:Nucleotides
keywords: DNA polymerase; Modified nucleotide; Reversible terminator; Sequencing-by-synthesis; Triphosphate