期刊
NANOTECHNOLOGY
卷 26, 期 8, 页码 -出版社
IOP PUBLISHING LTD
DOI: 10.1088/0957-4484/26/8/084002
关键词
DNA stretching; electroosmosis; single-molecule; ionic strength
资金
- European Research Council (European Commission's Seventh Framework Programme) [260884]
- European Research Council (ERC) [260884] Funding Source: European Research Council (ERC)
In nanopore sequencing, where single DNA strands are electrophoretically translocated through a nanopore and the resulting ionic signal is used to identify the four DNA bases, an enzyme has been used to ratchet the nucleic acid stepwise through the pore at a controlled speed. In this work, we investigated the ability of alpha-hemolysin nanopores to distinguish the four DNA bases under conditions that are compatible with the activity of DNA-handling enzymes. Our findings suggest that in immobilized strands, the applied potential exerts a force on DNA (similar to 10 pN at +160 mV) that increases the distance between nucleobases by about 2.2 angstrom V-1. The four nucleobases can be resolved over wide ranges of applied potentials (from +60 to +220 mV in 1 m KC1) and ionic strengths (from 200 mM KC1 to 1M KC1 at +160 mV) and nucleobase recognition can be improved when the ionic strength on the side of the DNA-handling enzyme is low, while the ionic strength on the opposite side is high.
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