DNA strands from denatured duplexes are translocated through engineered protein nanopores at alkaline pH.
Nanopores are under development for the detection of a variety of analytes and the investigation of chemical reactions at the single molecule level. In particular, the analysis of nucleic acid molecules is under intense investigation, including the development of systems for rapid, low-cost DNA sequencing. Here, we show that DNA can be translocated through an engineered alphaHL protein pore at pH 11.7, a value at which dsDNA is denatured. Therefore, the alphaHL pore is sufficiently stable to entertain the possibility of direct nanopore sequencing of genomic dsDNA samples, which are more readily obtained and handled than ssDNA.
Maglia
Giovanni
G
Department of Chemistry, University of Oxford, Oxford, OX1 3TA, United Kingdom.
Henricus
Marsiyana
M
Wyss
Romain
R
Li
Qiuhong
Q
Cheley
Stephen
S
Bayley
Hagan
H
D004247:DNA / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D006460:Hemolysin Proteins / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D049329:Nanostructures / Q000737:chemistry; D036103:Nanotechnology; D009691:Nucleic Acid Denaturation; D011506:Proteins / Q000737:chemistry; D017422:Sequence Analysis, DNA / Q000379:methods
D006460:Hemolysin Proteins; D011506:Proteins; D004247:DNA