title: 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.
authors.affiliation: Department of Physics, University of Washington, Seattle, Washington, United States of America.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Craig
authors.affiliation: Department of Physics, University of Washington, Seattle, Washington, United States of America.
authors.forename: Andrew H
authors.initials: AH
authors.lastname: Laszlo
authors.affiliation: Department of Physics, University of Washington, Seattle, Washington, United States of America.
authors.forename: Ian M
authors.initials: IM
authors.lastname: Derrington
authors.affiliation: Department of Physics, University of Washington, Seattle, Washington, United States of America.
authors.forename: Brian C
authors.initials: BC
authors.lastname: Ross
authors.affiliation: Department of Physics, University of Washington, Seattle, Washington, United States of America.
authors.forename: Henry
authors.initials: H
authors.lastname: Brinkerhoff
authors.affiliation: Department of Physics, University of Washington, Seattle, Washington, United States of America.
authors.forename: Ian C
authors.initials: IC
authors.lastname: Nova
authors.affiliation: Department of Physics, University of Washington, Seattle, Washington, United States of America.
authors.forename: Kenji
authors.initials: K
authors.lastname: Doering
authors.affiliation: Department of Physics, University of Washington, Seattle, Washington, United States of America.
authors.forename: Benjamin I
authors.initials: BI
authors.lastname: Tickman
authors.affiliation: Department of Physics, University of Washington, Seattle, Washington, United States of America.
authors.forename: Mark T
authors.initials: MT
authors.lastname: Svet
authors.affiliation: Department of Physics, University of Washington, Seattle, Washington, United States of America.
authors.forename: Jens H
authors.initials: JH
authors.lastname: Gundlach
mesh_terms: D017103:Bacillus Phages; D001426:Bacterial Proteins / Q000235:genetics; D004247:DNA / Q000032:analysis* / Q000235:genetics; D004265:DNA Helicases / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D003854:Deoxyribonucleotides / Q000032:analysis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005815:Genetic Code; D007477:Ions; D008051:Lipid Bilayers / Q000737:chemistry; D058608:Nanopores; D009711:Nucleotides / Q000032:analysis* / Q000235:genetics; D018272:Porins / Q000235:genetics*; D017422:Sequence Analysis, DNA; D019712:Thermococcus / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D003854:Deoxyribonucleotides; D007477:Ions; D008051:Lipid Bilayers; D009711:Nucleotides; D018272:Porins; C121838:mspA protein, Mycobacterium smegmatis; D004247:DNA; D004259:DNA-Directed DNA Polymerase; D004265:DNA Helicases