Direct Detection of Unnatural DNA Nucleotides dNaM and d5SICS using the MspA Nanopore

Malyshev et al. showed that the four-letter genetic code within a living organism could be expanded to include the unnatural DNA bases dNaM and d5SICS. However, verification and detection of these unnatural bases in DNA requires new sequencing techniques. Here we provide proof of concept detection o...

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Autores principales: Craig, Jonathan M., Laszlo, Andrew H., Derrington, Ian M., Ross, Brian C., Brinkerhoff, Henry, Nova, Ian C., Doering, Kenji, Tickman, Benjamin I., Svet, Mark T., Gundlach, Jens H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4654578/
https://www.ncbi.nlm.nih.gov/pubmed/26588074
http://dx.doi.org/10.1371/journal.pone.0143253
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author Craig, Jonathan M.
Laszlo, Andrew H.
Derrington, Ian M.
Ross, Brian C.
Brinkerhoff, Henry
Nova, Ian C.
Doering, Kenji
Tickman, Benjamin I.
Svet, Mark T.
Gundlach, Jens H.
author_facet Craig, Jonathan M.
Laszlo, Andrew H.
Derrington, Ian M.
Ross, Brian C.
Brinkerhoff, Henry
Nova, Ian C.
Doering, Kenji
Tickman, Benjamin I.
Svet, Mark T.
Gundlach, Jens H.
author_sort Craig, Jonathan M.
collection PubMed
description Malyshev et al. showed that the four-letter genetic code within a living organism could be expanded to include the unnatural DNA bases dNaM and d5SICS. However, verification and detection of these unnatural bases in DNA requires new sequencing techniques. Here we provide proof of concept detection of dNaM and d5SICS in DNA oligomers via nanopore sequencing using the nanopore MspA. We find that both phi29 DNA polymerase and Hel308 helicase are capable of controlling the motion of DNA containing dNaM and d5SICS through the pore and that single reads are sufficient to detect the presence and location of dNaM and d5SICS within single molecules.
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spelling pubmed-46545782015-11-25 Direct Detection of Unnatural DNA Nucleotides dNaM and d5SICS using the MspA Nanopore Craig, Jonathan M. Laszlo, Andrew H. Derrington, Ian M. Ross, Brian C. Brinkerhoff, Henry Nova, Ian C. Doering, Kenji Tickman, Benjamin I. Svet, Mark T. Gundlach, Jens H. PLoS One Research Article Malyshev et al. showed that the four-letter genetic code within a living organism could be expanded to include the unnatural DNA bases dNaM and d5SICS. However, verification and detection of these unnatural bases in DNA requires new sequencing techniques. Here we provide proof of concept detection of dNaM and d5SICS in DNA oligomers via nanopore sequencing using the nanopore MspA. We find that both phi29 DNA polymerase and Hel308 helicase are capable of controlling the motion of DNA containing dNaM and d5SICS through the pore and that single reads are sufficient to detect the presence and location of dNaM and d5SICS within single molecules. Public Library of Science 2015-11-20 /pmc/articles/PMC4654578/ /pubmed/26588074 http://dx.doi.org/10.1371/journal.pone.0143253 Text en © 2015 Craig et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Craig, Jonathan M.
Laszlo, Andrew H.
Derrington, Ian M.
Ross, Brian C.
Brinkerhoff, Henry
Nova, Ian C.
Doering, Kenji
Tickman, Benjamin I.
Svet, Mark T.
Gundlach, Jens H.
Direct Detection of Unnatural DNA Nucleotides dNaM and d5SICS using the MspA Nanopore
title Direct Detection of Unnatural DNA Nucleotides dNaM and d5SICS using the MspA Nanopore
title_full Direct Detection of Unnatural DNA Nucleotides dNaM and d5SICS using the MspA Nanopore
title_fullStr Direct Detection of Unnatural DNA Nucleotides dNaM and d5SICS using the MspA Nanopore
title_full_unstemmed Direct Detection of Unnatural DNA Nucleotides dNaM and d5SICS using the MspA Nanopore
title_short Direct Detection of Unnatural DNA Nucleotides dNaM and d5SICS using the MspA Nanopore
title_sort direct detection of unnatural dna nucleotides dnam and d5sics using the mspa nanopore
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4654578/
https://www.ncbi.nlm.nih.gov/pubmed/26588074
http://dx.doi.org/10.1371/journal.pone.0143253
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