impact of thawing
Wednesday 7 December 2011
Biobanks of fresh, unfixed human tissue represent a valuable source for gene expression analysis in translational research and molecular pathology.
Minimal RNA degradation was detected after 30 minutes of thawing in unfixed samples. This degradation was accompanied by relevant changes in gene expression for FOS and BCL2 at 45 minutes.
Modified primer design or the use of different housekeeping genes could not rectify the changes for FOS.
Repetitive thawing cycles had similar effects on RNA integrity.
The incubation of the tissue in RNAlater efficiently prevented RNA degradation.
In conclusion, degradation of RNA in frozen tissue occurs first after several minutes of thawing.
Already minimal decrease in RNA quality may result in significant changes in gene expression patterns in clinical tissue samples.
Impact of thawing on RNA integrity and gene expression analysis in fresh frozen tissue. Botling J, Edlund K, Segersten U, Tahmasebpoor S, Engström M, Sundström M, Malmström PU, Micke P. Diagn Mol Pathol. 2009 Mar;18(1):44-52. PMID: #19214109#