Discrimination of oligonucleotides of different lengths with a wild-type aerolysin nanopore

Nat Nanotechnol. 2016 Aug;11(8):713-8. doi: 10.1038/nnano.2016.66. Epub 2016 Apr 25.

Abstract

Protein nanopores offer an inexpensive, label-free method of analysing single oligonucleotides. The sensitivity of the approach is largely determined by the characteristics of the pore-forming protein employed, and typically relies on nanopores that have been chemically modified or incorporate molecular motors. Effective, high-resolution discrimination of oligonucleotides using wild-type biological nanopores remains difficult to achieve. Here, we show that a wild-type aerolysin nanopore can resolve individual short oligonucleotides that are 2 to 10 bases long. The sensing capabilities are attributed to the geometry of aerolysin and the electrostatic interactions between the nanopore and the oligonucleotides. We also show that the wild-type aerolysin nanopores can distinguish individual oligonucleotides from mixtures and can monitor the stepwise cleavage of oligonucleotides by exonuclease I.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Toxins / chemistry*
  • Hydrogen-Ion Concentration
  • Nanopores*
  • Nanotechnology / methods*
  • Oligonucleotides / analysis
  • Oligonucleotides / chemistry*
  • Oligonucleotides / isolation & purification*
  • Pore Forming Cytotoxic Proteins / chemistry*
  • Static Electricity

Substances

  • Bacterial Toxins
  • Oligonucleotides
  • Pore Forming Cytotoxic Proteins
  • aerolysin