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Horm Metab Res 2006; 38(1): 8-11
DOI: 10.1055/s-2006-924968
Original Basic
© Georg Thieme Verlag KG Stuttgart · New York

Real-time PCR: Housekeeping Genes in the INS-1E β-cell Line

K.  Smidt1 , L.  Wogensen2 , B.  Brock1 , O.  Schmitz1, 3 , J.  Rungby1, 3
  • 1Department of Clinical Pharmacology, University of Aarhus, Denmark
  • 2Research Laboratory for Biochemical Pathology, Aarhus University Hospital, NBG, Denmark
  • 3Departments of Endocrinology and Metabolism C+M, Aarhus University Hospital, Aarhus, Denmark
Further Information

Publication History

Received 23 June 2005

Accepted after revision 8 September 2005

Publication Date:
13 February 2006 (online)

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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.

Key words

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

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Jørgen Rungby

Department of Clinical Pharmacology · University of Aarhus · Denmark ·

Phone: +45 8942 1708

Fax: +45 8612 8804

Email: jr@farm.au.dk