Data analysis of translocation events in nanopore experiments

Abstract

Nanopores have become important tools for single molecule experiments, where information about the properties of DNA/RNA or proteins is inferred from current pulses elicited by individual molecules as they traverse a single pore. However, because of necessary electronic filters employed in the measurement technique, the extraction of meaningful information from short pulses is limited. This restricts the use of nanopores for the investigation of small molecules which cross the pore rapidly. Here we present a method which significantly improves the accuracy of the analysis of noise-filtered current pulses. We introduce improved criteria to measure the pulse width and propose a method to evaluate the pulse height from the falling edge of the pulse, which renders the identification of a pulse plateau unnecessary. The new methods are compared to conventional routines and validated by analyzing representative current pulses as well as experimental protein translocation data. It is demonstrated that the pulse properties can be recovered with satisfying accuracy beyond the usual limitations of Bessel filters, i.e., from pulses featuring a width of merely 0.3f(c)(-1) (f(c) being the filter cutoff frequency).