Isolation and detection methods of plant miRNAs

Pedro Fernando Vera-Hernández, Stefan de Folter, Flor de Fátima Rosas-Cárdenas

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

Abstract

Small RNAs (sRNAs) are RNAs of low abundance in organisms. Among sRNAs, miRNAs are included and represent approximately 10% of the total number of sRNAs. The isolation of sRNAs is critical for miRNA detection and analysis. The precipitation of low-molecular-weight (LMW) RNAs from total RNA extracts has allowed enrichment of sRNAs. Here, we describe a simple method to isolate sRNAs from different plant species. The main advantage of this method is that it does not need first an extraction of total RNA and it is not based on TRIzol® reagent. This method has been successfully used for miRNA analyses by Northern blot assay and RT-qPCR (these techniques are as well described in this chapter), as well as sRNA library preparation.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages109-120
Number of pages12
DOIs
StatePublished - 2019

Publication series

NameMethods in Molecular Biology
Volume1932
ISSN (Print)1064-3745

Keywords

  • Plant miRNAs
  • miRNA analysis
  • miRNA detection
  • sRNA extraction

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