RNA Stability in Human Liver: Comparison of Different Processing Times, Temperatures and Methods

• Published in: Molecular biotechnology 2013, Vol.53 (1), p.1-8
• Main Authors: Lee, Serene M. L., Schelcher, Celine, Gashi, Sevdije, Schreiber, Stefanie, Thasler, Reinhard M. K., Jauch, Karl-Walter, Thasler, Wolfgang E.
• Format: Article
• Language: English
• Subjects: Biochemistry; Biological Techniques; Biotechnology; Blood; Cell Biology; Chemistry; Chemistry and Materials Science; DNA, Complementary - genetics; DNA, Complementary - isolation & purification; Exons; Fos protein; Freezing; Gene Expression; Human Genetics; Humans; Ice; Liver; Liver - metabolism; Molecular modelling; Myc protein; Polymerase chain reaction; Protein Science; Quantitative genetics; Real-Time Polymerase Chain Reaction; Ribonucleic acid; RNA; RNA - genetics; RNA - isolation & purification; RNA Stability - genetics; Sampling; Specimen Handling - methods; Surgery; Temperature effects; Time Factors; Tissue Banks
• ISSN: 1073-6085; 1559-0305
• DOI: 10.1007/s12033-011-9493-4

The accuracy of information garnered by real-time quantitative polymerase chain reaction (RT-qPCR), an important technology for elucidating molecular mechanisms of disease, is dependent on tissue quality. Thus, this study aimed to determine the effects of intra-operative manipulation, extended processing times, different temperatures or storage in RNAlater on RNA quality in liver samples for tissue banking. Liver samples, flash-frozen or in RNAlater, were collected over a time course (during surgery before blood arrest up to 1 day after surgery) with samples kept either at room temperature (RT) or on ice. This study showed that at the longest time-point at RT, the RNA quality decreased significantly by 20%. However, relative gene expressions of FOS , GUSB , MYC , HIF1 α and GFER were in general not significantly different when the time-points were compared. In conclusion, samples should be kept on ice during processing, and either RNAlater or snap-freezing should be utilised for storage. Further, intra-operative manipulation and extended postoperative processing time generally does not change relative gene expression levels for the 5 genes studied, making such sampling suitable for RT-qPCR analysis. Thus, if relative gene expression of a gene of interest is stable, these guidelines will lead to increased accrual of samples to the tissue bank.