Real-time PCR: housekeeping genes in the INS-1E beta-cell line.
Investigation of gene expression is a developing area with several methods available. One method is quantitative PCR. A major pitfall in quantitative PCR is the normalisation procedure of the gene expression. Many experiments include a housekeeping gene, some use RNA concentration, and others use a geometric mean of several internal, stably expressed genes. This study demonstrates that real-time-PCR results differ with varying housekeeping genes and analysis protocols when applied to insulin-secreting INS-1E cells derived from the pancreas and stimulated by DEDTC (diethyldithiocarbamate, a zinc chelator) and GLP-1.
Department of Clinical Pharmacology, University of Aarhus, Denmark.
K
K
Smidt
L
L
Wogensen
B
B
Brock
O
O
Schmitz
J
J
Rungby
D000818:Animals; D002460:Cell Line; D002614:Chelating Agents / Q000494:pharmacology; D004050:Ditiocarb / Q000031:analogs & derivatives / Q000494:pharmacology; D020869:Gene Expression Profiling / Q000379:methods / Q000592:standards; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000502:physiology*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000592:standards
D002614:Chelating Agents; D007328:Insulin; D004050:Ditiocarb; C039353:dihydroxyethyldithiocarbamate