TaqMan PCR assay in the control of RNA normalization in human post-mortem brain tissue

Neurochemistry International
Marta BarrachinaIsidro Ferrer

Abstract

The brain tissue obtained after death is subjected to several circumstances that can affect RNA integrity. The present study has been directed to reveal possible pitfalls and to control RNA normalization in post-mortem samples in order to recognize the limitations and minimize errors when using TaqMan PCR technology. This has been carried out in samples of the frontal cortex in a series of control and diseased cases covering Parkinson's disease, dementia with Lewy bodies pure form and common form, and Alzheimer's disease. Special attention has been paid to the value of the agonal state, post-mortem delay and pH of the nervous tissue as approximate predictors of the quality of RNA, as well as to the use of the Bioanalyzer to confirm RNA preservation. In addition, since possible disease-modified mRNAs have to be normalized with ideal unaltered RNAs, TaqMan human endogenous control plates have been used to determine the endogenous control most appropriate for the study. beta-glucuronidase (GUS) and beta-actin were good endogenous controls because their expression levels showed a small variation across a representative number of control and pathological cases. RNA stability was also analysed in a paradigm mimicking cumulative delay...Continue Reading

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Citations

Mar 28, 2008·Acta Neuropathologica·Jeanne E BellHans Kretzschmar
Aug 7, 2012·Acta Neuropathologica·Pascal F DurrenbergerRichard Reynolds
Aug 13, 2010·Cell and Tissue Banking·Alex C BirdsillThomas G Beach
Sep 17, 2011·Cell and Tissue Banking·Marta BarrachinaIsidre Ferrer
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Aug 20, 2010·Forensic Science International : Synergy·Hitoshi MaedaTomomi Michiue
Dec 17, 2009·Journal of Neuropathology and Experimental Neurology·Pascal F DurrenbergerRichard Reynolds
May 10, 2013·Journal of Neuropathology and Experimental Neurology·Paula Garcia-EsparciaIsidre Ferrer
Mar 11, 2015·Journal of Neuropathology and Experimental Neurology·Irene López-GonzálezIsidre Ferrer
Jun 1, 2007·Forensic Science International. Genetics·Marielle HeinrichUlrike Schmidt
Mar 1, 2007·Forensic Science International. Genetics·M Bauer
Jan 20, 2015·Neuropathology and Applied Neurobiology·Paula Garcia-EsparciaIsidre Ferrer
Jun 24, 2009·Journal of Neurochemistry·Edna GrünblattPeter Riederer
Feb 5, 2008·Brain Pathology·José L AlbasanzMairena Martín
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Sep 8, 2007·The American Journal of Pathology·Marta BarrachinaIsidre Ferrer
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Aug 25, 2015·Neural Plasticity·Sara BerjaouiIsidre Ferrer
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Dec 17, 2011·Brain Research Bulletin·Patrick A Lewis, Mark R Cookson
Jan 25, 2017·Neuropathology and Applied Neurobiology·P Andrés-BenitoI Ferrer
May 15, 2018·Frontiers in Aging Neuroscience·Paula Garcia-EsparciaIsidro Ferrer
Feb 6, 2017·Journal of Neuropathology and Experimental Neurology·Irene López-GonzálezIsidro Ferrer
May 23, 2018·Journal of Neuropathology and Experimental Neurology·Pol Andrés-BenitoIsidro Ferrer

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