|
Taqman
- Human GAPDH Protocol
Primers and Probe
GAPDH Forward Primer
5'-GAAGGTGAAGGTCGGAGTC-3'
GAPDH Reverse Primer
5'-GAAGATGGTGATGGGATTTC-3'
GAPDH Probe
JOE-CAAGCTTCCCGTTCTCAGCC-TAMRA
Master Mix Recipe
(use 5痞 of RT reaction)
|
GAPDH Master Mix Components
|
Volume (無) per Reaction
|
Concentration in Final Reaction
|
| H2O |
22.75 無
|
---
|
| 10X TaqMan buffer A |
5 無
|
1 X
|
| 25 mM MgCl2 |
11 無
|
5.5 mM
|
| 2% gelatin |
1.25 無
|
0.05%
|
| 25 mM dNTPs |
1 無
|
500 然
|
| 5 然 GAPDH Probe |
1 無
|
100 nM
|
| 10 然 GAPDH Forward Primer |
1 無
|
200 nM
|
| 10 然 GAPDH Reverse Primer |
1 無
|
200 nM
|
| AmpErase UNG |
0.5 無
|
0.01 U/無
|
| AmpliTaq Gold |
0.5 無
|
0.05 U/無
|
Thermal Cycler Conditions
|
Temperature
|
Time
|
Cycles
|
| 50蚓 |
2 minutes |
|
| 95蚓 |
10 minutes |
|
| 95蚓 |
15 seconds |
40 cycles
|
| 60蚓 |
1 minute |
| 
