title: Validation of RNA extraction procedures focused on micro RNA expression analysis.
abstract: The sampling procedure is a crucial step in every kind of experiment. This is also true in gene expression profiling experiments, where high quality and sufficient quantity of extracted RNA plays a significant role in the experimental outcome. We have compared five different RNA extraction protocols from peripheral blood/PBMCs with the aim to define the most suitable method for the miRNA expression profiling experiments. Convincing results in terms of highest quantity and quality were obtained by the TRIzol-chloroform extraction method. The total RNA obtained using this method contained the highest portion of good-quality miRNA molecules, which was also confirmed by gene-specific real-time PCR experiments.
authors.affiliation: Department of Experimental and Clinical Rheumatology, Institute of Rheumatology, Prague, Czech Republic.
authors.forename: M
authors.initials: M
authors.lastname: Remáková
authors.forename: M
authors.initials: M
authors.lastname: Škoda
authors.forename: M
authors.initials: M
authors.lastname: Faustová
authors.forename: J
authors.initials: J
authors.lastname: Vencovský
authors.forename: P
authors.initials: P
authors.lastname: Novota
mesh_terms: D000818:Animals; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods*; D015203:Reproducibility of Results
chemical_list: D035683:MicroRNAs