Translocation of RecA-coated double-stranded DNA through solid-state nanopores.
We report translocation of double-stranded DNA (dsDNA) molecules that are coated with RecA protein through solid-state nanopores. Translocation measurements show current-blockade events with a wide variety in time duration (10-4-10-1 s) and conductance blockade values (3-14 nS). Large blockades (11.4+/-0.7 nS) are identified as being caused by translocations of RecA-dsDNA filaments. We confirm these results through a variety of methods, including changing molecular length and using an optical tweezer system to deliver bead-functionalized molecules to the nanopore. We further distinguish two different regimes of translocation: a low-voltage regime (<150 mV) in which the event rate increases exponentially with voltage, and a high-voltage regime in which it remains constant. Our results open possibilities for a variety of future experiments with (partly) protein-coated DNA molecules, which is interesting for both fundamental science and genomic screening applications.
Smeets
R M M
RM
KaVli Institute of Nanoscience, Delft University of Technology, Lorentzweg 1, 2628 CJ Delft, The Netherlands.
Kowalczyk
S W
SW
Hall
A R
AR
Dekker
N H
NH
Dekker
C
C
D004247:DNA / Q000032:analysis / Q000378:metabolism*; D008567:Membranes, Artificial; D049329:Nanostructures / Q000737:chemistry*; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods*; D010316:Particle Size; D011938:Rec A Recombinases / Q000378:metabolism*; D013499:Surface Properties; D013997:Time Factors
D008567:Membranes, Artificial; D004247:DNA; D011938:Rec A Recombinases