Denaturing reversed phase liquid chromatographic separation of non-coding ribonucleic acids on macro-porous polystyrene-divinylbenzene resins

J Chromatogr A. 2013 Oct 18;1312:87-92. doi: 10.1016/j.chroma.2013.09.021. Epub 2013 Sep 9.

Abstract

The ability of denaturing ion-paired reversed phase LC to separate RNA was assessed using macro-porous polystyrene-divinylbenzene resins as the stationary phase. Using the three stationary phases with different pore size and a mobile phase containing phosphate, we separated RNAs of 20-8000 nucleotides with extremely high sensitivity, e.g., 50pg for an RNA 20 nucleotides in length, S/N=5. The method was used to separate non-coding RNAs obtained from biological sources and is suited for use with direct MS-based chemical characterization.

Keywords: Denaturing reversed phase high-performance liquid chromatography; Macro-porous polystyrene-divinylbenzene resin; Mass spectrometry; RNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Chromatography, Reverse-Phase / instrumentation
  • Chromatography, Reverse-Phase / methods*
  • HEK293 Cells
  • Humans
  • Molecular Sequence Data
  • Molecular Weight
  • Polystyrenes / chemistry*
  • RNA, Untranslated / analysis
  • RNA, Untranslated / chemistry
  • RNA, Untranslated / isolation & purification
  • Sensitivity and Specificity

Substances

  • Polystyrenes
  • RNA, Untranslated
  • divinylbenzene-polystyrene copolymer