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

Autor: Jonathan M Craig, Andrew H Laszlo, Ian M Derrington, Brian C Ross, Henry Brinkerhoff, Ian C Nova, Kenji Doering, Benjamin I Tickman, Mark T Svet, Jens H Gundlach
Jazyk: angličtina
Rok vydání: 2015
Předmět:
Zdroj: PLoS ONE, Vol 10, Iss 11, p e0143253 (2015)
Druh dokumentu: article
ISSN: 1932-6203
DOI: 10.1371/journal.pone.0143253
Popis: 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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