DNA sequencing by synthesis using 3'-O-azidomethyl nucleotide reversible terminators and surface-enhanced Raman spectroscopic detection.

ABSTRACT: As an alternative to fluorescence-based DNA sequencing by synthesis (SBS), we report here an approach using an azido moiety (N₃) that has an intense, narrow and unique Raman shift at 2125 cm^-1, where virtually all biological molecules are transparent, as a label for SBS. We first demonstrated that the four 3'-O-azidomethyl nucleotide reversible terminators (3'-O-azidomethyl-dNTPs) displayed surface enhanced Raman scattering (SERS) at 2125 cm^-1. Using these 4 nucleotide analogues as substrates, we then performed a complete 4-step SBS reaction. We used SERS to monitor the appearance of the azide-specific Raman peak at 2125 cm^-1 as a result of polymerase extension by a single 3'-O-azidomethyl-dNTP into the growing DNA strand and

SUBMITTER: Palla M 

PROVIDER: S-EPMC4226440 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature