title: Effects of post-mortem and physical degradation on RNA integrity and quality.
abstract: The precision and reliability of quantitative nucleic acid analysis depends on the quality of the sample analyzed and the integrity of the nucleic acids. The integrity of RNA is currently primarily assessed by the analysis of ribosomal RNA, which is the by far dominant species. The extrapolation of these results to mRNAs and microRNAs, which are structurally quite different, is questionable. Here we show that ribosomal and some nucleolar and mitochondrial RNAs, are highly resistant to naturally occurring post-mortem degradation, while mRNAs, although showing substantial internal variability, are generally much more prone to nucleolytic degradation. In contrast, all types of RNA show the same sensitivity to heat. Using qPCR assays targeting different regions of mRNA molecules, we find no support for 5' or 3' preferentiality upon post-mortem degradation.
authors.affiliation: Laboratory of Gene Expression, Institute of Biotechnology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague, Czech Republic; Charles University in Prague, Faculty of Science, Department of Cell Biology, Vinicna 7, 128 43, Prague, Czech Republic.
authors.forename: Monika
authors.initials: M
authors.lastname: Sidova
authors.affiliation: Laboratory of Gene Expression, Institute of Biotechnology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague, Czech Republic.
authors.forename: Silvie
authors.initials: S
authors.lastname: Tomankova
authors.affiliation: Laboratory of Gene Expression, Institute of Biotechnology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague, Czech Republic.
authors.forename: Pavel
authors.initials: P
authors.lastname: Abaffy
authors.affiliation: Laboratory of Gene Expression, Institute of Biotechnology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague, Czech Republic; TATAA Biocenter AB, Odinsgatan 28, 411 03 Göteborg, Sweden.
authors.forename: Mikael
authors.initials: M
authors.lastname: Kubista
authors.affiliation: Laboratory of Gene Expression, Institute of Biotechnology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague, Czech Republic.
authors.forename: Radek
authors.initials: R
authors.lastname: Sindelka
keywords: Degradation; RNA integrity; RNA quality; RQI; RT-qPCR