title: How the RNA isolation method can affect microRNA microarray results.
abstract: The quality of RNA is crucial in gene expression experiments. RNA degradation interferes in the measurement of gene expression, and in this context, microRNA quantification can lead to an incorrect estimation. In the present study, two different RNA isolation methods were used to perform microRNA microarray analysis on porcine brain tissue. One method is a phenol-guanidine isothiocyanate-based procedure that permits isolation of total RNA. The second method, miRVana™ microRNA isolation, is column based and recovers the small RNA fraction alone. We found that microarray analyses give different results that depend on the RNA fraction used, in particular because some microRNAs appear very sensitive to the RNA isolation method. We conclude that precautions need to be taken when comparing microarray studies based on RNA isolated with different methods.
authors.affiliation: Department of Basic Animal and Veterinary Sciences, Section of Genetics and Bioinformatics, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Podolska
authors.forename: Bogumil
authors.initials: B
authors.lastname: Kaczkowski
authors.forename: Thomas
authors.initials: T
authors.lastname: Litman
authors.forename: Merete
authors.initials: M
authors.lastname: Fredholm
authors.forename: Susanna
authors.initials: S
authors.lastname: Cirera
mesh_terms: D000818:Animals; D020869:Gene Expression Profiling / Q000379:methods; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D020871:RNA Stability; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017423:Sequence Analysis, RNA / Q000379:methods; D013552:Swine
chemical_list: D035683:MicroRNAs; D011933:Reagent Kits, Diagnostic; D012313:RNA