title: Determining abundance of microRNAs and other small RNAs in legumes.
abstract: High-throughput sequencing techniques have been applied to the discovery of several different regulatory small RNAs, including siRNAs and microRNAs in different plant species. Their subsequent characterization demands the use of sensitive and quantitative methods for their detection. Here we describe the use of northern blot and quantitative PCR for these purposes. We highlight the advantages and shortcomings of each method and offer a detailed description of those techniques that best work in our hands, in particular having in mind their use for the study of several small RNAs in a given sample. We enumerate relevant alternatives as well as cautionary comments in cases where we have detected potential difficulties.
authors.affiliation: Departamento de Biología Molecular de Plantas, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca, Mexico.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Contreras-Cubas
authors.forename: Alejandra A
authors.initials: AA
authors.lastname: Covarrubias
authors.forename: José Luis
authors.initials: JL
authors.lastname: Reyes
mesh_terms: D015152:Blotting, Northern / Q000379:methods; D007887:Fabaceae / Q000235:genetics*; D035683:MicroRNAs / Q000235:genetics*; D018749:RNA, Plant; D058727:RNA, Small Untranslated / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods
chemical_list: D035683:MicroRNAs; D018749:RNA, Plant; D058727:RNA, Small Untranslated