Journal
BIOCHEMISTRY AND CELL BIOLOGY
Volume 82, Issue 3, Pages 407-412Publisher
CANADIAN SCIENCE PUBLISHING, NRC RESEARCH PRESS
DOI: 10.1139/O04-005
Keywords
DNA unzipping; bacterial nanopores; DNA transport; single-molecule detection; DNA mismatch
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A 50-base Guide strand was synthesized that consisted of a central 10-base probe sequence flanked by two tracts of 20 adenine residues. Target sequences of 10 bases containing up to three mismatches were prepared and hybridized to the Guide strand in 1 M KCl. The transport of these constructs through single alpha-hemolysin pores was analysed by measuring the current blockade as a function of time. Complementary dsDNA takes significantly longer (840 +/- 60 mus) to pass through the pore than a sequence of the same length containing a single (590 +/- 45 mus) and a double (270 +/- 50 mus) mismatch. Constructs involving three mismatches were indistinguishable from Guide ssDNA transport (120 +/- 30 mus). The results suggest that dsDNA must unzip as it is transported through the nanopore. Duplexes containing mismatches unzip more quickly and can be distinguished from those with perfect complementarity.
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