Direct Detection of Unnatural DNA Nucleotides dNaM and d5SICS using the MspA Nanopore.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 26588074.
- Also identified by DOI 10.1371/journal.pone.0143253 and PMC identifier 4654578.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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.
Medical subject headings
- DNA
- Deoxyribonucleotides
- Nanopores
- Nucleotides
- Porins