Controlling DNA capture and propagation through artificial nanopores.
Electrophorescing biopolymers across nanopores modulates the ionic current through the pore, revealing the polymer's diameter, length, and conformation. The rapidity of polymer translocation ( approximately 30,000 bp/ms) in this geometry greatly limits the information that can be obtained for each base. Here we show that the translocation speed of lambda-DNA through artificial nanopores can be reduced using optical tweezers. DNAs coupled to optically trapped beads were presented to nanopores. DNAs initially placed up to several micrometers from the pore could be captured. Subsequently, the optical tweezers reduced translocation speeds to 150 bp/ms, about 200-fold slower than free DNA. Moreover, the optical tweezers allowed us to "floss" single polymers back and forth through the pore. The combination of controlled sample presentation, greatly slowed translocation speeds, and repeated electrophoresis of single DNAs removes several barriers to using artificial nanopores for sequencing, haplotyping, and characterization of protein-DNA interactions.
Trepagnier
Eliane H
EH
Biophysics Graduate Group, University of California, Berkeley, Berkeley, California, USA.
Radenovic
Aleksandra
A
Sivak
David
D
Geissler
Phillip
P
Liphardt
Jan
J
D004247:DNA / Q000737:chemistry* / Q000528:radiation effects*; D004058:Diffusion; D004574:Electromagnetic Fields; D018274:Electroporation / Q000379:methods*; D008567:Membranes, Artificial; D009038:Motion; D049329:Nanostructures / Q000737:chemistry* / Q000528:radiation effects*; D052898:Optical Tweezers; D016062:Porosity
D008567:Membranes, Artificial; D004247:DNA