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Vol 46(2012) N 1 p. 153-160; A. Birerdinc1,2, R. Mehta1,2, R. Alhussain1,3, A. Afendi2,4, V. Chandhoke1,2, Z. Younossi1,2,4, A.V. Baranova1,2,5* Selection of reliable reference genes for qRT-PCR analysis in human non-cancerous gastric tissue 1School of Systems Biology, George Mason University, Fairfax, USA2Translational Research Institute, Inova Health System, Falls Church, USA 3Biology Department, College of Science, George Mason University, Fairfax, USA 4Center for Liver Diseases, Inova Fairfax Hospital, Falls Church, USA 5Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Moscow, 115478 Russia *aancha@gmail.com Received - 2011-04-04; Accepted - 2011-06-07 Real-time PCR (qRT-PCR) is the standard method for studying changes in relative gene expression in complex diseases like obesity and gastritis. However, variations in amount of starting material, enzymatic efficiency and presence of amplification inhibitors can lead to quantification errors. Hence, the need for accurate data normalization is vital. Among several known strategies for data normalization, the use of reference genes as an internal control is the most common approach. Human gastric tissue has been the least investigated for stability of reference gene expression. In this study, three popular algorithms, GeNorm, NormFinder and BestKeeper were used to evaluate the reference gene stability. Conclusion: HPRT1 and GAPDH are the best performing pair of reference genes for qRT-PCR profiling experiments involving non - malignant gastric tissue samples. qRT-PCR, housekeeping genes, endogenous control, stomach, obesity |
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