Relative Quantification - CRN2M
Relative Quantification - CRN2M
Relative Quantification - CRN2M
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Comparative Ct Method: an Example Using the Four<br />
Samples<br />
∆Rn<br />
t=0<br />
∆Rn<br />
t=12 h<br />
Ct=15<br />
Ct=35<br />
Cycles<br />
Ct=15<br />
Ct=30<br />
Cycles<br />
∆Rn<br />
t=24 h ∆Rn<br />
t=48 h<br />
Ct=9 Ct=24 Cycles<br />
Ct=14<br />
Endogenous control<br />
Target gene<br />
25<br />
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Ct=34<br />
Cycles<br />
Comparative Ct Method Calculation Steps<br />
step 1: Normalization to endogenous control<br />
Ct Target gene – Ct Endogenous control = ∆Ct<br />
Do both for calibrator and samples<br />
step 2: Normalization to calibrator sample<br />
∆Ct Sample – ∆Ct Calibrator = ∆∆Ct<br />
step 3: use the formula<br />
2<br />
-∆∆Ct<br />
26<br />
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