Virtual terminator nucleotides for next-generation DNA sequencing

Abstract

We synthesized reversible terminators with tethered inhibitors for next generation sequencing. These were efficiently incorporated with high fidelity while preventing incorporation of additional nucleotides and were used to sequence canine bacterial artificial chromosomes in a single-molecule system that provided even coverage for over 99% of the region sequenced. This single-molecule approach generated high quality sequence data without the need for target amplification and thus avoided concomitant biases.