摘要 |
The present invention relates to a DNA sequencing method using a nucleoside triphosphate with a fluorescent blocking group on its 3'-OH end as a reversible terminator. Further, the present invention relates to sequencing-by-synthesis method using the mono-modified reversible terminator (MRT), the novel nucleotide monomer having a reversible fluorescent blocking group removable chemically or enzymatically at its 3'-OH end. The sequencing method of the present invention facilitates sequencing of bases inserted by terminating extension of a nucleotide chain by the nucleotide monomer and then detecting fluorescence signal from 3'-OH end. At this time, after analyzing the fluorescence signal, the blocking group conjugated to the 3'-OH end can be effectively removed, indicating that a free 3'-OH functional group can be successfully restored, so that the next monomer insertion is possible, making continuous sequencing possible.
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