Immuno-northern blotting: Detection of modified RNA using gel separation and antibodies to modified nucleosides

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2 Citations (Scopus)

Abstract

Immuno-northern blotting is a method for detecting modified RNAs using gel separation and specific antibodies to modified nucleosides. This method was developed by combining two commonly used molecular biology techniques: western blotting and northern blotting. In this method, urea-polyacrylamide (or agarose) gel-separated RNAs are transferred to positively charged nylon membrane and then immune detection is performed with specific antibodies to modified nucleosides: such as 1-methyladenosine, N6-methyladenosine, and pseudouridine. This highly sensitive and relatively simple method, which uses widely available laboratory equipment, enables small laboratories to compare the abundance of modified nucleic acids across samples.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages179-187
Number of pages9
DOIs
Publication statusPublished - 2019

Publication series

NameMethods in Molecular Biology
Volume1870
ISSN (Print)1064-3745

Keywords

  • 1-Methyladenosine
  • Modified nucleotide
  • N6-Methyladenosine
  • Pseudouridine
  • RNA modification
  • Small RNAs
  • tRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Mishima, E., & Abe, T. (2019). Immuno-northern blotting: Detection of modified RNA using gel separation and antibodies to modified nucleosides. In Methods in Molecular Biology (pp. 179-187). (Methods in Molecular Biology; Vol. 1870). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8808-2_13