title: TaqMan™ fluorogenic detection system to analyze gene transcription in autopsy material.
abstract: Real-time polymerase chain reaction using a TaqMan fluorogenic detection system is a simple and sensitive assay for quantitative analysis of gene transcription. This method is of potential usefulness in quantifying mRNA of a target gene in autopsy material that has undergone only a small amount of postmortem degradation. The TaqMan fluorogenic detection system can monitor PCR in real time using a dual-labeled fluorogenic hybridization probe (TaqMan probe) and a polymerase with 5'-3' exonuclease activity. The procedures of the quantitative reverse transcription polymerase chain reaction are as follows: RNA is extracted from autopsy material and used to synthesize cDNA by an RT reaction, and the target of interest is amplified and detected by the real-time PCR. The absolute amount of target mRNA in the sample is then determined relative to a standard curve. This chapter describes the methodology of the TaqMan fluorogenic detection system in handling autopsy material in the gene transcription assay.
authors.affiliation: Department of Legal Medicine, Osaka City University Medical School, Asahimachi 1-4-3, Osaka, 545-8585, Japan, kaori@m.u-tokyo.ac.jp.
authors.forename: Kaori
authors.initials: K
authors.lastname: Shintani-Ishida
authors.forename: Bao-Li
authors.initials: BL
authors.lastname: Zhu
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Maeda
mesh_terms: D001344:Autopsy; D015342:DNA Probes / Q000737:chemistry / Q000235:genetics*; D005456:Fluorescent Dyes / Q000737:chemistry; D020869:Gene Expression Profiling; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction; D019914:Taq Polymerase; D014158:Transcription, Genetic; D059467:Transcriptome
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes; D019914:Taq Polymerase