title: Quantitative analysis of gene expression relative to 18S rRNA in carcinoma samples using the LightCycler instrument and a SYBR GreenI-based assay: determining FAT10 mRNA levels in hepatocellular carcinoma.
abstract: Due to the fact that mutations and up- or downregulation of genes can lead to the development of cancer, quantitative comparison of relative gene expression in healthy and cancerous tissue can gain valuable insights into tumorigenesis. While the semi-quantitative DNA microarrays are being used to identify differentially expressed genes on a genomic scale, real-time RT-PCR provides a powerful tool for quantitative measurement of gene expression. Presently, it is the most sensitive method available. Here we describe in detail a SYBR GreenI-based assay using the LightCycler instrument to measure the levels of mRNA for the ubiquitin-like protein FAT10 relative to 18S rRNA in human hepatocellular carcinoma tissue. This method can be easily adapted to any tissue (human or mouse, rat, etc.) and any gene.
authors.affiliation: Department of Biology, University of Konstanz Universitätsstr, Konstanz, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Lukasiak
authors.forename: Kai
authors.initials: K
authors.lastname: Breuhahn
authors.forename: Claudia
authors.initials: C
authors.lastname: Schiller
authors.forename: Gunter
authors.initials: G
authors.lastname: Schmidtke
authors.forename: Marcus
authors.initials: M
authors.lastname: Groettrup
mesh_terms: D052160:Benzothiazoles; D006528:Carcinoma, Hepatocellular / Q000235:genetics*; D003959:Diamines; D015870:Gene Expression; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics*; D009930:Organic Chemicals; D011804:Quinolines; D012333:RNA, Messenger / Q000032:analysis*; D012337:RNA, Ribosomal, 18S / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014452:Ubiquitins / Q000235:genetics*
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; D012333:RNA, Messenger; D012337:RNA, Ribosomal, 18S; C109184:UBD protein, human; D014452:Ubiquitins; C098022:SYBR Green I