Title |
Nucleotide Discrimination with DNA Immobilized in the MspA Nanopore
|
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Published in |
PLOS ONE, October 2011
|
DOI | 10.1371/journal.pone.0025723 |
Pubmed ID | |
Authors |
Elizabeth A. Manrao, Ian M. Derrington, Mikhail Pavlenok, Michael Niederweis, Jens H. Gundlach |
Abstract |
Nanopore sequencing has the potential to become a fast and low-cost DNA sequencing platform. An ionic current passing through a small pore would directly map the sequence of single stranded DNA (ssDNA) driven through the constriction. The pore protein, MspA, derived from Mycobacterium smegmatis, has a short and narrow channel constriction ideally suited for nanopore sequencing. To study MspA's ability to resolve nucleotides, we held ssDNA within the pore using a biotin-NeutrAvidin complex. We show that homopolymers of adenine, cytosine, thymine, and guanine in MspA exhibit much larger current differences than in α-hemolysin. Additionally, methylated cytosine is distinguishable from unmethylated cytosine. We establish that single nucleotide substitutions within homopolymer ssDNA can be detected when held in MspA's constriction. Using genomic single nucleotide polymorphisms, we demonstrate that single nucleotides within random DNA can be identified. Our results indicate that MspA has high signal-to-noise ratio and the single nucleotide sensitivity desired for nanopore sequencing devices. |
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Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 3 | 1% |
United Kingdom | 1 | <1% |
Korea, Republic of | 1 | <1% |
Germany | 1 | <1% |
Unknown | 197 | 97% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 56 | 28% |
Researcher | 31 | 15% |
Student > Master | 29 | 14% |
Student > Bachelor | 27 | 13% |
Student > Doctoral Student | 9 | 4% |
Other | 24 | 12% |
Unknown | 27 | 13% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 46 | 23% |
Agricultural and Biological Sciences | 39 | 19% |
Engineering | 21 | 10% |
Physics and Astronomy | 18 | 9% |
Chemistry | 18 | 9% |
Other | 27 | 13% |
Unknown | 34 | 17% |