Sizing DNA using a nanometer-diameter pore

被引:243
作者
Heng, JB [1 ]
Ho, C [1 ]
Kim, T [1 ]
Timp, R [1 ]
Aksimentiev, A [1 ]
Grinkova, YV [1 ]
Sligar, S [1 ]
Schulten, K [1 ]
Timp, G [1 ]
机构
[1] Univ Illinois, Urbana, IL 61801 USA
关键词
D O I
10.1529/biophysj.104.041814
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Each species from bacteria to human has a distinct genetic fingerprint. Therefore, a mechanism that detects a single molecule of DNA represents the ultimate analytical tool. As a first step in the development of such a tool, we have explored using a nanometer-diameter pore, sputtered in a nanometer-thick inorganic membrane with a tightly focused electron beam, as a transducer that detects single molecules of DNA and produces an electrical signature of the structure. When an electric field is applied across the membrane, a DNA molecule immersed in electrolyte is attracted to the pore, blocks the current through it, and eventually translocates across the membrane as verified unequivocally by gel electrophoresis. The relationship between DNA translocation and blocking current has been established through molecular dynamics simulations. By measuring the duration and magnitude of the blocking current transient, we can discriminate single-stranded from double-stranded DNA and resolve the length of the polymer.
引用
收藏
页码:2905 / 2911
页数:7
相关论文
共 35 条
[1]   Microsecond time-scale discrimination among polycytidylic acid, polyadenylic acid, and polyuridylic acid as homopolymers or as segments within single RNA molecules [J].
Akeson, M ;
Branton, D ;
Kasianowicz, JJ ;
Brandin, E ;
Deamer, DW .
BIOPHYSICAL JOURNAL, 1999, 77 (06) :3227-3233
[2]   Microscopic kinetics of DNA translocation through synthetic nanopores [J].
Aksimentiev, A ;
Heng, JB ;
Timp, G ;
Schulten, K .
BIOPHYSICAL JOURNAL, 2004, 87 (03) :2086-2097
[3]   DIRECT NANOMETER SCALE PATTERNING OF SIO2 WITH ELECTRON-BEAM IRRADIATION [J].
ALLEE, DR ;
UMBACH, CP ;
BROERS, AN .
JOURNAL OF VACUUM SCIENCE & TECHNOLOGY B, 1991, 9 (06) :2838-2841
[4]   Resistive-pulse sensing - From microbes to molecules [J].
Bayley, H ;
Martin, CR .
CHEMICAL REVIEWS, 2000, 100 (07) :2575-2594
[5]   Stochastic sensors inspired by biology [J].
Bayley, H ;
Cremer, PS .
NATURE, 2001, 413 (6852) :226-230
[6]  
BROERS AN, 1978, ELECT MICROSCOPY, V3
[7]   A 2ND GENERATION FORCE-FIELD FOR THE SIMULATION OF PROTEINS, NUCLEIC-ACIDS, AND ORGANIC-MOLECULES [J].
CORNELL, WD ;
CIEPLAK, P ;
BAYLY, CI ;
GOULD, IR ;
MERZ, KM ;
FERGUSON, DM ;
SPELLMEYER, DC ;
FOX, T ;
CALDWELL, JW ;
KOLLMAN, PA .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1995, 117 (19) :5179-5197
[8]   Characterization of nucleic acids by nanopore analysis [J].
Deamer, DW ;
Branton, D .
ACCOUNTS OF CHEMICAL RESEARCH, 2002, 35 (10) :817-825
[9]   THE UBIQUITOUS PRESENCE OF CHANNELS WITH WIDE LUMENS AND THEIR GATING BY VOLTAGE [J].
FINKELSTEIN, A .
ANNALS OF THE NEW YORK ACADEMY OF SCIENCES, 1985, 456 :26-32
[10]   Identification and characterization of a cell membrane nucleic acid channel [J].
Hanss, B ;
Leal-Pinto, E ;
Bruggeman, LA ;
Copeland, TD ;
Klotman, PE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (04) :1921-1926