Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes
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Cites background or methods from "Accurate normalization of real-time..."
...Unfortunately, literature shows that as of yet, no such housekeeping gene has been found (1–7), and a recent study touching on the subject even stated that such a gene does not exist (8)....
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...We then ranked the candidates using another recently published strategy to identify stably expressed genes; in the remainder of this article, this strategy is called the pairwise comparison approach (8)....
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...In our pursuit of suitable normalization genes, we first turned to previously reported approaches for evaluating normalization gene candidates (8, 13)....
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Cites methods from "Accurate normalization of real-time..."
...For a subsequent data processing, the CP datasets can be imported into analysis software tools such as REST (Pfaffl et al. 2002), GeNorm ( Vandesompele et al. 2002 ) or Q-Gene (Muller et al. 2002)....
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...The earlier presented GeNorm software (Vandesompele et al. 2002) is restricted to the HKG analysis only, whereas, in BestKeeper software, additionally up to ten TGs can be analysed....
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...For a subsequent data processing, the CP datasets can be imported into analysis software tools such as REST (Pfaffl et al. 2002), GeNorm (Vandesompele et al. 2002) or Q-Gene (Muller et al. 2002)....
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...…parameters of the quantification procedure (e.g. inhibitory factors of the tissue, integrity of the RNA, loading error, enzyme or primer performance, etc.) must be controlled, numerous standardisation methods have been proposed (Suzuki et al. 2000, Thellin et al. 1999, Vandesompele et al. 2002)....
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...inhibitory factors of the tissue, integrity of the RNA, loading error, enzyme or primer performance, etc.) must be controlled, numerous standardisation methods have been proposed (Suzuki et al. 2000, Thellin et al. 1999, Vandesompele et al. 2002 )....
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"Accurate normalization of real-time..." refers methods in this paper
...The average gene-specific variation (determined as coefficient of variation, in percent) for the three control genes with the smallest variation within each tissue panel after normalization with three different factors calculated as the geometric mean of the three control genes with the lowest (NF3(1-3)), highest (NF3(8-10)) and intermediate (NF3(6-8)) gene stability values (as determined by geNorm)....
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