A biophysicist’s guide to translocation of double-stranded nucleic acids through solid-state nanopores

Biosensing with solid-state nanopores (SSNPs) is a versatile technique suitable for the analysis of double-stranded DNA, RNA, and their hybrids, complementing the use of biological nanopores for single-stranded nucleic acids and proteins. In this review, we systematically explain how operational parameters - including bias voltage, pore geometry, and buffer properties - affect translocation of double-stranded nucleic acids through SSNPs. We focus on key performance metrics for resolution, characterised by translocation time and signal-to-noise ratio, and throughput, described by event frequency and unfolded fraction. Beyond summarizing empirical observations, we dissect the underlying physical mechanisms that mediate these dependencies, including electrophoretic driving, hydrodynamic friction, electro-osmotic flow, and entropic effects. By integrating experimental findings with physical models, we aim to provide a practical guide for optimizing and interpreting SSNP experiments. The concepts discussed are broadly transferable to other SSNP targets and nanoscale transport applications.