Nanopore-based identification of individual nucleotides for direct RNA sequencing

Nano Lett. 2013;13(12):6144-50. doi: 10.1021/nl403469r. Epub 2013 Nov 13.

Abstract

We describe a label-free ribobase identification method, which uses ionic current measurement to resolve ribonucleoside monophosphates or diphosphates in α-hemolysin protein nanopores containing amino-cyclodextrin adapters. The accuracy of base identification is further investigated through the use of a guanidino-modified adapter. On the basis of these findings, an exosequencing approach is envisioned in which a processive exoribonuclease (polynucleotide phosphorylase) presents sequentially cleaved ribonucleoside diphosphates to a nanopore.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Hemolysin Proteins / chemistry
  • Nanopores*
  • Nanostructures / chemistry
  • Nucleotides / chemistry
  • Nucleotides / isolation & purification*
  • Sequence Analysis, RNA*

Substances

  • Hemolysin Proteins
  • Nucleotides