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Forming an α-Hemolysin Nanopore for Single-Molecule Analysis

Protocol
First Online:
Part of the Methods in Molecular Biology™ book series (MIMB, volume 544)

Summary

Nanopore analysis of single molecules can be performed by measuring the modulation in ionic current passing through the nanopore while an individual biomolecule such as DNA or RNA is resident in, translocating through, or otherwise interacting with the pore. The corresponding current signature has been shown to reveal properties of the biomolecule and information on its interactions with the pore. The α-hemolysin nanopore remains the pore of choice, particularly for single-molecule analysis of nucleic acids, because of its internal dimensions, hydrophilicity, and low-noise characteristics. In this chapter we present a detailed protocol for forming a robust α-hemolysin nanopore (or multiple nanopores) for single-molecule analysis.

Key words

Nanopore α-Hemolysin, Lipid bilayer, Single-molecule analysis 
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Notes

Acknowledgments

We thank members of the University of British Columbia Applied Biophysics Laboratory, in particular, Vincent Tabard-Cossa and Dhruti Trivedi. Many thanks to Jonathan Nakane, Mark Akeson, and Carolina Tropini for their important contributions to this protocol. This work was supported by the US National Institutes of Health (NIH). Financial support for Matthew Wiggin was provided by the Natural Sciences and Engineering Research Council of Canada (NSERC).

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  1. 1.Department of Physics and AstronomyUniversity of British ColumbiaVancouverCanada

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