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: Contribution to journalArticle

4 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 identification and significance 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 efficient design for allowing the downstream evaluation of miRNA and protein expression in mitochondria.

Original languageEnglish (US)
Pages (from-to)9-22
Number of pages14
JournalMethods in Molecular Biology
Volume1241
DOIs
StatePublished - Jan 1 2015

Keywords

  • Differential centrifugation
  • MiRNA
  • Mitochondria
  • Protein
  • TRIzol

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