Novel 3′-O-Fluorescently Modified Nucleotides for Reversible Termination of DNA Synthesis

Life Sciences Research Division, Korea Institute of Science and Technology, Cheongryang, Seoul, Korea.
ChemBioChem (Impact Factor: 3.09). 01/2009; 11(1):75-8. DOI: 10.1002/cbic.200900632
Source: PubMed


(Chemical Equation Presented) Serving a double purpose: 3′-O-fluorophore-labeled dTTPs were synthesized and used as reversible terminators of DNA polymerization. Successful incorporation of nucleotides with a bulky 3′-O-fluorophore and detection of fluorescence for base calling can potentially be used to improve sequencing-by-synthesis technology.

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    • "Within the scope of this study, several 3′-blocking groups were investigated, including bulky esters[7] and ethers[8] with the label attached to the blocking group and small groups. Some examples are the 3′-O-(2-nitrobenzyl) group investigated by Metzker and co-workers[6] and Welch and Burgess,[8, 11] the 3′-O-allyl group reported by Metzker,[6] Ju,[12] and Kim,[13] or the 3′-O-azidomethyl group, which was used by Ju and co-workers[14, 15] and was also realised in a commercially available device, the Genome Analyzer developed by Illumina/Solexa.[16, 17] Other interesting groups are the 3′-O-NH2 group from Benner and co-workers,[18] the 3′-O-(2-cyanoethoxy)methyl group reported by us,[19] or some 3′-blocking groups removable under mild reducing or mild acidic conditions reported by Kwiatkowski.[20] "
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