RNA and microRNA Stability in PAXgene-Fixed Paraffin-Embedded Tissue Blocks After Seven Years' Storage.
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Accepted version
Author(s)
Type
Journal Article
Abstract
Objectives: To evaluate the stability of RNA and microRNA (miRNA) in PAXgene-fixed paraffin-embedded tissue blocks after 7 years' storage. Methods: RNA and miRNA were extracted from PAXgene-fixed paraffin-embedded (PFPE) blocks in 2009 then stored at -80°C. Seven years later, RNA and miRNA were again extracted from the same blocks. RNA and miRNA integrity in the 2009 and 2016 extractions were compared using RNA integrity number (RIN), paraffin-embedded RNA metric (PERM), reverse transcription polymerase chain reaction (RT-PCR) for different amplicon lengths, and quantitative RT-PCR (qRT-PCR) for three mRNA and three miRNA targets. Results: In PFPE blocks, mRNA was poorer in 2016 extractions compared to the 2009 extractions in all blocks and all assays applied, with transcripts degrading at different rates in the same blocks. For miRNA, qRT-PCR showed no statistically significant differences between 2009 and 2016 extractions. Conclusions: mRNA in PFPE tissue blocks degrades at room temperature storage over 7 years.
Date Issued
2018-04-05
Date Acceptance
2018-04-01
Citation
American Journal of Clinical Pathology, 2018, 149 (6), pp.536-547
ISSN
0002-9173
Publisher
Oxford University Press (OUP)
Start Page
536
End Page
547
Journal / Book Title
American Journal of Clinical Pathology
Volume
149
Issue
6
Copyright Statement
© American Society for Clinical Pathology, 2018. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com. This is a pre-copy-editing, author-produced version of an article accepted for publication in American Journal of Clinical Pathology, following peer review. The definitive publisher-authenticated version Ignacio Sanchez, Fay Betsou, Bénédicte Culot, Sonia Frasquilho, Siobhan C McKay, Stephanos Pericleous, Colleen Smith, Gerry Thomas, William Mathieson; RNA and microRNA Stability in PAXgene-Fixed Paraffin-Embedded Tissue Blocks After Seven Years’ Storage, American Journal of Clinical Pathology, Volume 149, Issue 6, 25 April 2018, Pages 536–547, is available online at: https://doi.org/10.1093/ajcp/aqy026
Sponsor
Imperial College Healthcare NHS Trust- BRC Funding
Imperial College Healthcare NHS Trust- BRC Funding
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/29659661
PII: 4962388
Grant Number
RDC01 79560
RDF03
Subjects
11 Medical And Health Sciences
Pathology
Publication Status
Published
Coverage Spatial
England
Date Publish Online
2018-04-05