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Evaluation of potential reference genes for qRT-PCR data normalization in HeLa cells |
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| Authors: | N. A. Krainova N. A. Khaustova D. S. Makeeva N. N. Fedotov E. A. Gudim E. A. Ryabenko M. U. Shkurnikov V. V. Galatenko D. A. Sakharov D. V. Maltseva |
| Affiliation: | 1. SRC Bioclinicum, Moscow, Russia 2. Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, Moscow, Russia 3. Faculty of Mechanics and Mathematics, Lomonosov Moscow State University, Moscow, Russia 4. Institute of General Pathology and Pathophysiology, Moscow, Russia |
| Abstract: | 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 (ΔCt, geNorm, qBase and Haller’s equivalence test) were applied for the reference genes identification in HeLa cell line which is one of the most popular cellular models. Expression stability of seven candidate genes (HPRT1, ACTB, GAPDH, RPS18, HSPC3, UBC and SDHA) was determined at standard conditions, under heat shock and during relaxation. The genes RPS18 and HSPC3 were chosen as reference after the combination of all the validation methods. |
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