Isolation of mitochondria from liver and extraction of total RNA and protein: Analyses of MicroRNA and protein expressions

Pedro M. Borralho, Clifford J. Steer, Cecília M P Rodrigues

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

Several studies have indicated the presence of microRNAs (miRNAs) within mitochondria, although the origin, as well as the biological function of these mitochondrially located miRNAs is largely unknown. The identifi cation and signifi cance of this subcellular localization is gaining increasing relevance to the pathogenesis of certain disease states. Here we describe the isolation of highly purifi ed mitochondria from rat liver by differential centrifugation, followed by RNAse A treatment to eliminate contaminating RNA. The coupled extraction of total RNA and protein is a more effi cient design for allowing the downstream evaluation of miRNA and protein expression in mitochondria.

Original languageEnglish (US)
Title of host publicationMitochondrial Regulation
Subtitle of host publicationMethods and Protocols
PublisherSpringer New York
Pages9-22
Number of pages14
ISBN (Electronic)9781493918751
ISBN (Print)9781493918744
DOIs
StatePublished - Oct 11 2014

Keywords

  • Differential centrifugation
  • miRNA
  • Mitochondria
  • Protein
  • TRIzol

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