eprintid: 19930
rev_number: 13
eprint_status: archive
userid: 1589
dir: disk0/00/01/99/30
datestamp: 2016-02-09 13:18:54
lastmod: 2024-03-10 04:00:27
status_changed: 2016-02-09 13:18:54
type: article
metadata_visibility: show
creators_name: Ibberson, David
creators_name: Benes, Vladimir
creators_name: Muckenthaler, Martina U.
creators_name: Castoldi, Mirco
title: RNA degradation compromises the reliability of microRNA expression profiling
subjects: 570
divisions: 718000
divisions: 850800
divisions: 910500
abstract: Background: MicroRNAs are small non-coding RNAs that post-transcriptionally regulate gene expression and their expression is frequently altered in human diseases, including cancer. To correlate clinically relevant parameters with microRNA expression, total RNA is frequently prepared from samples that were archived for various time periods in frozen tissue banks but, unfortunately, RNA integrity is not always preserved in these frozen tissues. Here, we investigate whether experimentally induced RNA degradation affects microRNA expression profiles.   Results: Tissue samples were maintained on ice for defined time periods prior to total RNA extraction, which resulted in different degrees of RNA degradation. MicroRNA expression was then analyzed by microarray analysis (miCHIP) or microRNA-specific real-time quantitative PCR (miQPCR). Our results demonstrate that the loss of RNA integrity leads to in unpredictability of microRNA expression profiles for both, array-based and miQPCR assays.   Conclusion: MicroRNA expression cannot be reliably profiled in degraded total RNA. For the profiling of microRNAs we recommend use of RNA samples with a RNA integrity number equal to or above seven.
date: 2009
publisher: BioMed Central
id_scheme: DOI
ppn_swb: 1655535730
own_urn: urn:nbn:de:bsz:16-heidok-199308
language: eng
bibsort: IBBERSONDARNADEGRADA2009
full_text_status: public
publication: BMC Biotechnology
volume: 9
number: 102
place_of_pub: London
pagerange: 1-9
issn: 1472-6750
citation:   Ibberson, David ; Benes, Vladimir ; Muckenthaler, Martina U. ; Castoldi, Mirco  (2009) RNA degradation compromises the reliability of microRNA expression profiling.  BMC Biotechnology, 9 (102).  pp. 1-9.  ISSN 1472-6750     
document_url: https://archiv.ub.uni-heidelberg.de/volltextserver/19930/1/12896_2009_Article_468.pdf