Capture and analysis of double‐stranded DNA with the α‐hemolysin nanopore: Fundamentals and applications

Abstract

Abstract The α‐hemolysin nanopore has attracted much attention as a tool for the single‐molecule analysis of DNA due to its potential as an ultra‐sensitive, specific, and label‐free sensing technique. The vast majority of DNA sensing research with the α‐hemolysin nanopore has focused on interrogating single‐stranded DNA. Nevertheless, the structure of the α‐hemolysin pore, specifically the circa 32.6 cubic nanometer vestibule, is of sufficient size for a short section of double‐stranded DNA (dsDNA) to reside before unzipping into its single‐stranded constituents. In this review, we describe past and current literature relating to the rich information that can be obtained from the interrogation of dsDNA while residing within the α‐hemolysin nanopore vestibule, and the subsequent voltage‐driven unzipping of the residing DNA into its single‐stranded constituents. Applications for dsDNA interrogation and unzipping that have been implemented include DNA sequencing, disease diagnosis, and the identification of epigenetic modifications.