Folia Biologica
Journal of Cellular and Molecular Biology, Charles University 

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Fol. Biol. 2003, 49, 211-216

https://doi.org/10.14712/fb2003049060211

The Use of Housekeeping Genes (HKG) as an Internal Control for the Detection of Gene Expression by Quantitative Real-Time RT-PCR

Veronika Ullmannová, C. Haškovec

Department of Molecular Genetics, Institute of Haematology and Blood Transfusion, Prague, Czech Republic

Received August 2003
Accepted October 2003

Quantitative real-time RT-PCR is a very useful technique for estimating gene expression at the mRNA level. The expression of a tested gene has to be compared with that of a control gene. Various housekeeping genes have been used as control genes in different systems. In our study we tested several housekeeping genes in the model of gene expression after induction of apoptosis and differentiation. The myeloid cell lines were incubated with phorbol esters, butyric acid and combination of TNFα and IFNγ to induce differentiation. Camptothecin was used for induction of apoptosis. Tested control genes included β2-microglobulin, GAPDH, 18S ribosomal RNA and abl. GAPDH was found to be the best control gene in the apoptotic system. Different control genes were suitable for different systems where differentiation or senescence was induced. Our results show that attention should be paid to the choice of an appropriate control gene of quantitative real-time RT-PCR for different experimental models and various experimental conditions.

Funding

This work was supported by grant CEZ L33/98:237360001 from the Ministry of Health of the Czech Republic.

References

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