Relative Quantification - CRN2M
Relative Quantification - CRN2M
Relative Quantification - CRN2M
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Selection of <strong>Quantification</strong> Method<br />
<br />
Comparative Ct method<br />
– Easy to use, even for large gene numbers<br />
– Good design process will result in very high assay<br />
efficiencies<br />
– Added amount of sample must not cause inhibition<br />
• The goal must be to optimize extraction, RT and PCR<br />
conditions to avoid inhibition<br />
9<br />
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Endogenous Control (EC)<br />
<br />
<br />
<br />
The perfect EC reflects the amount of cDNA per well<br />
EC has a constant expression level in all samples<br />
which are used in that study<br />
The EC normalizes for<br />
– RNA input measurement errors<br />
– RT efficiency variations<br />
10<br />
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