QPCR结果分析方法
Sample | Target Gene | CT | mean | Internal control | CT | mean | △CT | 2^-△CT | mean | 2^-△△CT |
C1 | Gene | 18.724 | 18.771 | internal control | 15.777 | 16.160 | 2.611 | 0.16372350 | 1.49837857 | 0.109 |
18.741 | 15.980 | |||||||||
18.847 | 16.723 | |||||||||
C2 | Gene | 16.751 | 16.668 | internal control | 17.199 | 17.426 | -0.758 | 1.69114457 | 1.129 | |
16.636 | 17.652 | |||||||||
16.616 | 17.426 | |||||||||
C3 | Gene | 17.510 | 17.817 | internal control | 17.449 | 17.150 | 0.667 | 0.62981499 | 0.420 | |
17.507 | 17.215 | |||||||||
18.433 | 16.785 | |||||||||
T1 | Gene | 18.232 | 18.198 | internal control | 16.402 | 16.511 | 1.687 | 0.31064384 | 0.207 | |
18.165 | 16.605 | |||||||||
18.197 | 16.527 | |||||||||
T2 | Gene | 17.276 | 17.284 | internal control | 18.910 | 18.996 | -1.712 | 3.27690383 | 2.187 | |
17.314 | 18.980 | |||||||||
17.261 | 19.098 | |||||||||
T3 | Gene | 17.348 | 17.498 | internal control | 19.168 | 19.043 | -1.545 | 2.91804069 | 1.947 | |
17.611 | 19.160 | |||||||||
17.535 | 18.801 |
(1)分别求出Gene和内参internal control的3个重复CT的平均值mean；
(2)求出△CT，即Gene的3个重复CT的均值减去内参internal control的3个重复CT的均值；
(3)求出每个样品的目的基因的变化量2^-△CT；
(4)求出对照和处理样本中2^-△CT的均值；
(5)计算每个样本中的Fold Change 或者 Relative Expression Level，即2^-△△CT，用每个样本的2^-△CT除以所有样本2^-△CT的均值。