Validating internal controls for quantitative plant gene expression

Therefore, the present study was conducted to identify suitable HKG(s) for normalization of gene expression in soybean.All ten HKGs displayed a wide range of Ct values in 21 sample pools, confirming that they were variably expressed.Furthermore, it is easy to use, allows high throughput production of data and eliminates the need for radioactive isotopes [].

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ΔCt approach and 'Stability index' were also used to analyze the expression stability in all 21 sample pools.Results from ΔCt approach and ge Norm indicated that Gene expression analysis is becoming much more prevalent since it promotes our understanding of biological processes.Compared with the traditional methods for transcript analysis including Northern blot, RNase protection analysis, ], which allows a straightforward comparison between RNAs that differ widely in their abundance.These genes are supposed to have constant expression levels between different samples, and are frequently used as a normalizer without proper validation.However, recent studies show that the transcriptional levels of these HKGs are not always stable, and that no single HKG has a constant level under all experimental conditions [].

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