Evaluation of potential reference genes for qRT-PCR data normalization in HeLa cells

Applied Biochemistry and Microbiology. 2013. Vol. 49. No. 9. P. 743–749.

Krainova N. A., Khaustova N. A., Makeeva D. S., Fedotov N. N., Gudim E. A., Ryabenko E A, Shkurnikov M. U., Galatenko V V, Sakharov D. A., Maltseva D. V.

Reference genes selection is one of the most important stages in qPCR data normalization when a problem of quantitative determination of gene expression is addressed. Stability of gene expression level in all experimental conditions is a basic criterion for the reference gene selection. Over the past decade a lot of publications concerning validation methods of suitable reference genes appeared. In this paper, the main approaches (∆Ct, geNorm, qBase and Haller’s equivalence test) were applied for the reference genes identi? fication in HeLa cell line which is one of the most popular cellular models. Expression stability of seven can? didate genes (HPRT1, ACTB, GAPDH, RPS18, HSPC3, UBC and SDHA) was determined at standard condi? tions, under heat shock and during relaxation. The genes RPS18 and HSPC3 were chosen as reference after the combination of all the validation methods

Ключевые слова: Reference genes, qRT–PCR, cell line HeLa, heat shock, expression stability, HSPA1A