| Autor: |
Elizabeth A. Manrao, Mikhail Pavlenok, Jens H. Gundlach, Michael Niederweis, Kyle W. Langford, Marcus D. Collins, Ian M. Derrington |
| Rok vydání: |
2010 |
| Předmět: |
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| Zdroj: |
Biophysical Journal. 98(3) |
| ISSN: |
0006-3495 |
| DOI: |
10.1016/j.bpj.2009.12.3267 |
| Popis: |
Biological nanopores are currently being investigated as a fast, low cost DNA sequencing platform. Single stranded DNA (ssDNA) is electrophoretically driven through a protein pore as the ionic current through the constriction is measured. The porin MspA of Mycobacterium smegmatis was mutated to produce a channel highly suitable for nanopore DNA sequencing. To study the resolution of the mutated porin MspA, we immobilize ssDNA within the pore using a streptavidin ‘anchor’. Each base, adenine, cytosine, thymine, and guanine, produces a distinct current signature when it is held within the nanopore. We examine homopolymer ssDNA with a single heteronucleotide substitution to determine the recognition site within MspA. Discrimination of a single base in a heteromeric ssDNA is performed with two single nucleotide polymorphisms (SNPs) where the polymorphism is positioned at the recognition site. Our results indicate that MspA has the ability to provide high-resolution single nucleotide discrimination.View Large Image | View Hi-Res Image | Download PowerPoint Slide |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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