title: Evaluation of reference genes and normalization strategy for quantitative real-time PCR in human pancreatic carcinoma.
abstract: Histologically verified pairs (n=10) of pancreatic tumors and non-neoplastic tissues were used for quantitative real-time PCR and the stability of 24 reference genes was analyzed with geNorm and NormFinder software. Raw C{q} values correlated with the degree of RNA degradation. This correlation was abolished by normalization to C{q} of 18S endogenous control gene. Both geNorm and NormFinder programs suggested EIF2B1, ELF1, MRPL19, and POP4 as the same most stable genes. We have thus identified suitable reference genes for future expression studies in pancreatic carcinoma. Normalization method reducing the effects of RNA degradation on the quality of results was also developed.
authors.affiliation: Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic.
authors.forename: Beatrice
authors.initials: B
authors.lastname: Mohelnikova-Duchonova
authors.forename: Martin
authors.initials: M
authors.lastname: Oliverius
authors.forename: Eva
authors.initials: E
authors.lastname: Honsova
authors.forename: Pavel
authors.initials: P
authors.lastname: Soucek
mesh_terms: D000368:Aged; D014408:Biomarkers, Tumor / Q000235:genetics*; D002277:Carcinoma / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D020717:Eukaryotic Initiation Factor-2B / Q000235:genetics; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D008297:Male; D008875:Middle Aged; D024101:Mitochondrial Proteins / Q000235:genetics; D009687:Nuclear Proteins / Q000235:genetics; D010179:Pancreas / Q000378:metabolism* / Q000473:pathology; D010190:Pancreatic Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D020871:RNA Stability; D060888:Real-Time Polymerase Chain Reaction; D012260:Ribonucleases / Q000235:genetics; D012261:Ribonucleoproteins / Q000235:genetics; D012269:Ribosomal Proteins / Q000235:genetics; D012984:Software; D014157:Transcription Factors / Q000235:genetics
chemical_list: D014408:Biomarkers, Tumor; C512897:ELF1 protein, human; D020717:Eukaryotic Initiation Factor-2B; C496590:MRPL19 protein, human; D024101:Mitochondrial Proteins; D009687:Nuclear Proteins; D012261:Ribonucleoproteins; D012269:Ribosomal Proteins; D014157:Transcription Factors; D012260:Ribonucleases; C105253:POP4 protein, human