Real-time PCR: housekeeping genes in the INS-1E beta-cell line

Horm Metab Res. 2006 Jan;38(1):8-11. doi: 10.1055/s-2006-924968.

Abstract

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.

MeSH terms

  • Animals
  • Cell Line
  • Chelating Agents / pharmacology
  • Ditiocarb / analogs & derivatives
  • Ditiocarb / pharmacology
  • Gene Expression Profiling* / methods
  • Gene Expression Profiling* / standards
  • Gene Expression Regulation / physiology*
  • Humans
  • Insulin / metabolism
  • Insulin Secretion
  • Insulin-Secreting Cells / cytology
  • Insulin-Secreting Cells / physiology*
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction* / methods
  • Reverse Transcriptase Polymerase Chain Reaction* / standards

Substances

  • Chelating Agents
  • Insulin
  • Ditiocarb
  • dihydroxyethyldithiocarbamate