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Fig. 2 | BMC Research Notes

Fig. 2

From: Reference genes for reverse transcription quantitative PCR in canine brain tissue

Fig. 2

Determination of the optimum number of reference genes for normalisation of RT-qPCR data. The GeNorm program calculates the normalisation factor (NF) for a given number of genes (n) and defines the pairwise variation (V) between the sequential normalisation factors NFn and NFn+1. For example V2/3 represents the variation in NFs using the two versus three most stable control genes. A low V value indicates little variation, implying that adding the extra gene has no significant effect. Pairwise variation (V) < 0.15 is regarded to result in valid normalisation. Using this cut-off value all evaluated brain regions require only two reference genes for accurate normalisation. Results are based on analysis of 7 brain areas of 7 dogs each: LF lobus frontalis, LP lobus parietalis, LT lobus temporalis, LO lobus occipitalis, H hippocampus, C cerebellum, T thalamus, All average of all areas

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