Data analysis of translocation events in nanopore experiments

Area of Science:

• Biophysics
• Analytical Chemistry
• Nanoscience

Background:

• Nanopores are crucial for single-molecule analysis of DNA, RNA, and proteins.
• Electronic filters in nanopore measurements limit the analysis of short current pulses from rapidly translocating molecules.
• Investigating small molecules using nanopores is challenging due to rapid transit times and signal limitations.

Purpose of the Study:

• To develop an improved method for analyzing noise-filtered current pulses in nanopore sensing.
• To enhance the accuracy of extracting information from short, filtered current pulses.
• To enable the investigation of small molecules using nanopore technology.

Main Methods:

• Introduction of refined criteria for measuring nanopore event pulse width.
• Proposal of a novel method to determine pulse height from the falling edge, eliminating the need for plateau identification.
• Validation of new methods against conventional routines using simulated and experimental protein translocation data.

Main Results:

• The new methods significantly improve the accuracy of analyzing filtered current pulses.
• Pulse properties can be recovered with high accuracy even for pulses narrower than typical Bessel filter limitations (down to 0.3f(c)(-1)).
• The technique successfully analyzes experimental protein translocation data, demonstrating practical applicability.

Conclusions:

• The developed method enhances the utility of nanopores for analyzing short current pulses.
• This advancement expands the scope of nanopore sensing to include the study of small, rapidly translocating molecules.
• Improved pulse analysis overcomes filter-induced limitations, advancing single-molecule biophysical measurements.