Author: Wang, Jiying; Wang, Yanping; Wang, Huaizhong; Hao, Xiaojing; Wu, Ying; Guo, Jianfeng
Title: Selection of Reference Genes for Gene Expression Studies in Porcine Whole Blood and Peripheral Blood Mononuclear Cells under Polyinosinic:Polycytidylic Acid Stimulation Document date: 2014_4_23
ID: xf4yy6i1_33
Snippet: It has been suggested that using the geometric mean of two or more reference genes for qRT-PCR studies might generate more reliable results (Vandesompele et al., 2002) . In order to determine the optimal number of required reference genes, we also performed pairwise variation (V n /V n+1 ) analysis by geNorm, as shown in Figure 3 . Based on the cut-off value of 0.15 proposed by geNorm program, four reference genes are necessary to obtain the lowe.....
Document: It has been suggested that using the geometric mean of two or more reference genes for qRT-PCR studies might generate more reliable results (Vandesompele et al., 2002) . In order to determine the optimal number of required reference genes, we also performed pairwise variation (V n /V n+1 ) analysis by geNorm, as shown in Figure 3 . Based on the cut-off value of 0.15 proposed by geNorm program, four reference genes are necessary to obtain the lowest control genes in the tested samples of both whole blood and PBMC. In practice, it is not always possible to measure multiple reference genes due to limited sample availability and cost. Additionally, the cut-off value of 0.15 is an empirical value and should not be taken as a too strict cut off value, as it is already suggested by the geNorm manual itself. Therefore, we recommend using the geometric mean of the first three stable genes, TBP, PPIA, and B2M, to normalize the gene expression of whole blood and PBMC with poly I:C stimulation.
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