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Toward Single Molecule DNA Sequencing: Direct Identification of Ribonucleoside and Deoxyribonucleoside 5‘-Monophosphates by Using an Engineered Protein Nanopore Equipped with a Molecular Adapter
Abstract
Individual nucleic acid molecules might be sequenced by the identification of nucleoside 5'-monophosphates as they are released by processive exonucleases. Here, we show that single molecule detection with a modified protein nanopore can be used to identify ribonucleoside and 2'-deoxyribonucleoside 5'-monophosphates, thereby taking a step along this path. Distinct levels of current block are observed for each of the four members of a set of nucleoside 5'-monophosphates when the molecules bind within a mutant alpha-hemolysin pore, (M113R)(7), equipped with the molecular adapter heptakis-(6-deoxy-6-amino)-beta-cyclodextrin. While our results compare favorably with alternative approaches, further work will be required to improve the accuracy of identification of the nucleic acid bases, to feed each released nucleotide into the pore, and to ensure that every nucleotide is captured by the adapter.
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Cited by
Sequencing technologies and genome sequencing
Pareek, C. S., Smoczynski, R., & Tretyn, A. (2011) · Journal of Applied Genetics · doi:10.1007/s13353-011-0057-x · Cited by 868
Single-nucleotide discrimination in immobilized DNA oligonucleotides with a biological nanopore
Stoddart, D., Heron, A. J., Mikhailova, E., Maglia, G., & Bayley, H. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0901054106 · Cited by 482
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Metadata
- Authors
- Astier, Y., Braha, O., & Bayley, H.
- Venue
- Journal of the American Chemical Society
- Publisher
- American Chemical Society (ACS)
- Published
- 2006-01-12
- Volume
- 128
- Issue
- 5
- Pages
- 1705-1710
- ISSN
- 0002-7863 (ISSN-L)
- 1520-5126
- OpenAlex
- W2051815061
- Cited by
- 315
- References
- 0
- FWCI
- 18.28
- Type
- journal article
- Language
- en
- Open access
- closed
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- Astier, Y., Braha, O., & Bayley, H. (2006). Toward Single Molecule DNA Sequencing: Direct Identification of Ribonucleoside and Deoxyribonucleoside 5‘-Monophosphates by Using an Engineered Protein Nanopore Equipped with a Molecular Adapter. Journal of the American Chemical Society. https://doi.org/10.1021/ja057123%2B
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