Isolation and Detection Methods of Plant miRNAs
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.
Key wordssRNA extraction Plant miRNAs miRNA analysis miRNA detection
We thank the Mexican National Council of Science and Technology (CONACyT) for the fellowship of PPVH. This work was financed by CONACyT grant CB-2013-221522 and SIP grants 20170477 and 20180545. Work in the SDF laboratory was financed by the CONACyT grants CB-2012-177739 and FC-2015-2/1061.
- 3.Chappell L, Baulcombe D, Molnár A (2006) Isolation and Cloning of Small RNAs from virus-infected Plants. Curr Protoc Microbiol 16:16H.2Google Scholar