An Integrated System for DNA Sequencing by Synthesis Using Novel Nucleotide Analogues

摘要

Tnhe Human Genome Project has concluded, but its suc-ncessful completion has increased, rather than decreased,nthe need for high-throughput DNA sequencing technologies.nThe possibility of clinically screening a full genome for annindividual’s mutations offers tremendous benefits, both fornpursuing personalized medicine and for uncovering thengenomic contributions to diseases. The Sanger sequencingnmethod, although enormously productive for more than 30nyears, requires an electrophoretic separation step that, unfor-ntunately, remains a key technical obstacle for achieving eco-nnomically acceptable full-genome results. Alternative sequencing approaches thus focus on innovations that can reduce costs.nThe DNA sequencing by synthesis (SBS) approach has shown great promise as a new sequencing platform, with par-nticular progress reported recently. The general fluorescent SBS approach involves (i) incorporation of nucleotide analogs bear-ning fluorescent reporters, (ii) identification of the incorporated nucleotide by its fluorescent emissions, and (iii) cleavage ofnthe fluorophore, along with the reinitiation of the polymerase reaction for continuing sequence determination. In this Account,nwe review the construction of a DNA-immobilized chip and the development of novel nucleotide reporters for the SBSnsequencing platform.