Identification of Valid Endogenous Control Genes for Determining Gene Expression in Human Glioma using the Roche RealTime ready Human Reference Gene Panel
The authors found that in general glioblastoma exhibited the highest expression levels and largest variability of the candidate genes, whereas normal brain showed the lowest level. In each of the tumor subgroups and across these groups, the NormFinder analyses identified a large number of genes suitable for normalization. However, this number was drastically reduced after inclusion of normal brain into the analyses: it was expected that only GAPDH, IPO8, RPL13A, SDHA, and TBP would not be differentially expressed; statistical analysis indicated favorable stability values for all of these genes, TBP and IPO8 being the most stable. These 5 genes represent different physiological pathways and may be regarded as universal reference genes applicable for accurate normalization of gene expression in human astrocytomas of different grades (WHO Grades II-IV) alone and compared with normal brain, thereby enabling longitudinally designed studies (e.g., in astrocytoma before and after malignant transformation). The results show that the Roche RealTime ready Human Reference Gene Panel is a perfect tool for analyzing a wide variety of different genes in order to identify suitable reference genes for further relative quantification experiments.
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