Correcting False Gene Expression Measurements From Degraded RNA Using RTQ-PCRPort, Matthias MD* †; Schmelz, Hans Ulrich MD* ‡; Stassen, Tanja§; Müeller, Kerstin*; Stockinger, Marcus MD*; Obermair, Richard*; Abend, Michael MD, PhD*Author Information *Bundeswehr Institute of Radiobiology, Munich †Department of Hematology and Oncology, Hannover Medical School ‡Department of Urology, Federal Armed Forces Hospital, Koblenz §Institute of Veterinary Pathology, Ludwig-Maximillian-University, Munich, Germany Supported by the German Ministry of Defense. Reprints: Hans Ulrich Schmelz, MD, Federal Armed Forces Institute of Radiobiology, Ernst-von-Bergmann-Kaserne, Neuherbergstr. 11, D-80937 Munich, Germany (e-mail: [email protected]). Competing interests statement: We declare that we have no significant competing financial, professional, or personal interests that might have influenced the performance or presentation of the work described in this manuscript. Diagnostic Molecular Pathology: March 2007 - Volume 16 - Issue 1 - pp 38-49 doi: 10.1097/01.pdm.0000213472.70054.94 Buy Metrics Abstract This paper describes a method allowing correcting false gene expression measured on highly degraded RNA using real-time quantitative reverse transcription-polymerase chain reaction (RTQ-PCR). RNA was isolated from different models (in vitro cell lines, in vivo models of human and dog) and different tissue types. In vitro RNA degradation and modeling of in vivo degradation were applied on intact and degraded total RNA. Gene expression (eg, Bcl-2, GAPDH, PGK, PSME3, RAB2, BAX) was measured using RTQ-PCR. 18S rRNA proved to be the most constant house-keeping gene. Less than 10-fold degraded RNA can be quantified correctly when using 18S rRNA for normalization purposes. Higher-fold degraded RNA can be quantified correctly up to a precision that is comparable to RTQ-PCR measurements on intact RNA when simulating the RNA-species and tissue-specific degradation kinetic. © 2007 Lippincott Williams & Wilkins, Inc.