Relative Quantification - CRN2M
Relative Quantification - CRN2M
Relative Quantification - CRN2M
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Validation Experiment using Comparative Ct Method<br />
35<br />
30 Target gene: s = - 3.35<br />
Ct<br />
25<br />
20<br />
15<br />
EC: s = - 3.39<br />
10<br />
0 2 4 6 8 10<br />
Log (amount of cDNA)<br />
• Ct remains constant!<br />
• Optimal slope is – 3.3 to –3.4<br />
3<br />
¢¡¤£ £ ¥§¦©¨ ¨ <br />
What if the Slopes are not Equal <br />
Ct<br />
35<br />
30<br />
25<br />
20<br />
15<br />
Endogenous<br />
control<br />
s= -3.31<br />
Target gene<br />
s = -4.03<br />
Change your target assay !<br />
10<br />
0 2 4 6 8 10<br />
Log (amount of cDNA)<br />
4<br />
¢¡¤£ £ ¥§¦©¨ ¨
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