Detection of RNA in Ribonucleoprotein Complexes by Blue Native Northern Blotting

Methods Mol Biol. 2021:2170:45-51. doi: 10.1007/978-1-0716-0743-5_4.

Abstract

Northern blotting is a classical technique that allows the detection of specific nucleic acids using radioactive or non-radioactive probes. Normally, nucleic acids are denatured and separated by agarose or polyacrylamide gel electrophoresis and transferred and fixed to a membrane prior to detection. Here, we describe a method to analyze specific RNA in native ribonucleoprotein complexes using blue native PAGE with subsequent northern blotting, crosslinking of RNA onto a suitable membrane, and detection using non-radioactive probes.

Keywords: Native northern blot; Non-radioactive nucleic acid detection; RNA-binding proteins; Ribonucleoprotein complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Northern / methods*
  • Nucleic Acids / chemistry
  • Nucleic Acids / metabolism
  • RNA / chemistry*
  • RNA-Binding Proteins / chemistry*
  • RNA-Binding Proteins / metabolism*
  • Ribonucleoproteins / chemistry*
  • Ribonucleoproteins / metabolism*

Substances

  • Nucleic Acids
  • RNA-Binding Proteins
  • Ribonucleoproteins
  • RNA