Fast, cost-effective and flexible DNA sequencing by roll-to-roll fluidics.

Abstract

Next-generation sequencing (NGS) technologies have achieved remarkable success in both biological research and clinical applications. However, in recent years, performance improvements have slowed due to fundamental limitations imposed by Poiseuille fluid dynamics in flow cells, which we overcome using Couette flow. Here we show NGS by roll-to-roll fluidics (r2r-fl), a cost-effective approach compatible with flexible biochip sizes. r2r-fl is a practical implementation of plane Couette flow, with up to 85-fold lower reagent consumption (US$0.16 per gigabase pair), rinsing times under 2 s and a reduction in paired-end 100-base pair sequencing turnaround from days to less than 12 h. The method maintains over 99.9% precision and 99.3% sensitivity of single nucleotide polymorphisms in the human genome, 99.9% mapping rate for Escherichia coli, and minimal nucleotide substitutions, deletions or insertions in the severe acute respiratory syndrome coronavirus 2 alpha strain. By lowering cost and time, r2r-fl enables rapid, scalable NGS for pathogen detection, cancer diagnostics and genetic disease profiling.