Rapid nanopore discrimination between single polynucleotide molecules.
A variety of different DNA polymers were electrophoretically driven through the nanopore of an alpha-hemolysin channel in a lipid bilayer. Single-channel recording of the translocation duration and current flow during traversal of individual polynucleotides yielded a unique pattern of events for each of the several polymers tested. Statistical data derived from this pattern of events demonstrate that in several cases a nanopore can distinguish between polynucleotides of similar length and composition that differ only in sequence. Studies of temperature effects on the translocation process show that translocation duration scales as approximately T(-2). A strong correlation exists between the temperature dependence of the event characteristics and the tendency of some polymers to form secondary structure. Because nanopores can rapidly discriminate and characterize unlabeled DNA molecules at low copy number, refinements of the experimental approach demonstrated here could eventually provide a low-cost high-throughput method of analyzing DNA polynucleotides.
Meller
A
A
Department of Molecular and Cellular Biology, Division of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA.
Nivon
L
L
Brandin
E
E
Golovchenko
J
J
Branton
D
D
D001427:Bacterial Toxins / Q000737:chemistry*; D001482:Base Composition; D001671:Biochemistry / Q000379:methods*; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000302:isolation & purification*; D006460:Hemolysin Proteins / Q000737:chemistry*; D008051:Lipid Bilayers; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D010713:Phosphatidylcholines / Q000737:chemistry*; D011089:Polydeoxyribonucleotides / Q000737:chemistry / Q000302:isolation & purification*; D013696:Temperature
D001427:Bacterial Toxins; D004277:DNA, Single-Stranded; D006460:Hemolysin Proteins; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D011089:Polydeoxyribonucleotides; C021091:staphylococcal alpha-toxin; C021016:1,2-diphytanoylphosphatidylcholine