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

ABSTRACT: 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.

SUBMITTER: Craig JM

PROVIDER: S-EPMC4654578 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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

Craig Jonathan M JM Laszlo Andrew H AH Derrington Ian M IM Ross Brian C BC Brinkerhoff Henry H Nova Ian C IC Doering Kenji K Tickman Benjamin I BI Svet Mark T MT Gundlach Jens H JH

PloS one 20151120 11

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 d ...[more]

PMID: 26588074

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