4.2 Article

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

Journal

HORMONE AND METABOLIC RESEARCH
Volume 38, Issue 1, Pages 8-11

Publisher

GEORG THIEME VERLAG KG
DOI: 10.1055/s-2006-924968

Keywords

DEDTC; GLP-1; UBC-7; HPRT; cyclophilin A; ZnT-3; ZnT-8

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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-1 E cells derived from the pancreas and stimulated by DEDTC (diethyldithiocarbamate, a zinc chelator) and GLP-1.

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