Interfacing solid‐state nanopores with gel media to slow DNA translocations. Issue 15 (3rd June 2015)

• Record Type: Journal Article
• Authors: Waugh, Matthew; Carlsen, Autumn; Sean, David; Slater, Gary W.; Briggs, Kyle; Kwok, Harold; Tabard‐Cossa, Vincent; Lapizco‐Encinas, B. H.; Hayes, M. A.
• Is Part Of: Electrophoresis. Volume 36:Issue 15(2015)
• Page Start: 1759
• Page End: 1767
• Publication Date: 2015-06-03
• Subjects: Electrophoresis -- Periodicals; Electrophoresis -- Periodicals; 541.372
• DOI: 10.1002/elps.201400488
• Languages: English
• ISSNs: 0173-0835

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <p>We demonstrate the ability to slow DNA translocations through solid‐state nanopores by interfacing the <italic>trans</italic> side of the membrane with gel media. In this work, we focus on two reptation regimes: when the DNA molecule is flexible on the length scale of a gel pore, and when the DNA behaves as persistent segments in tight gel pores. The first regime is investigated using agarose gels, which produce a very wide distribution of translocation times for 5 kbp dsDNA fragments, spanning over three orders of magnitude. The second regime is attained with polyacrylamide gels, which can maintain a tight spread and produce a shift in the distribution of the translocation times by an order of magnitude for 100 bp dsDNA fragments, if intermolecular crowding on the <italic>trans</italic> side is avoided. While previous approaches have proven successful at slowing DNA passage, they have generally been detrimental to the S/N, capture rate, or experimental simplicity. These results establish that by controlling the regime of DNA movement exiting a nanopore interfaced with a gel medium, it is possible to address the issue of rapid biomolecule translocations through nanopores—presently one of the largest hurdles facing nanopore‐based analysis—without affecting the signal quality or capture efficiency.</p> </abstract>