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Infrared Laser Heating Applied to Nanopore Sensing for DNA Duplex Analysis

Angevine, Christopher E., Seashols-Williams, Sarah J., Reiner, Joseph E.
Analytical chemistry 2016 v.88 no.5 pp. 2645-2651
DNA, absorption, analytical methods, fluorescence, heat, hemolysins, melting, nanopores, temperature
Temperature studies coupled with resistive-pulse nanopore sensing enable the quantification of a variety of important thermodynamic properties at the single-molecule limit. Previous demonstrations of nanopore sensing with temperature control have utilized bulk chamber heating methodologies. This approach makes it difficult to rapidly change temperatures and enable optical access for other analytical techniques (i.e., single-molecule fluorescence). To address these issues, researchers have explored laser-based methodologies through either direct infrared (IR) absorption or plasmonic assisted heating. In this paper, we demonstrate the use of IR-based direct absorption heating with the DNA sensing capabilities of a biological nanopore. The IR heating enables rapid changes of the temperature in and around an α-hemolysin pore, and we use this to explore melting properties for short (≤50 bp) double-stranded DNA homopolymers. We also demonstrate that the IR heating enables one to measure the percentage of different-sized DNA molecules in a binary mixture.