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HOME > Protocols > Molecular Biology > Real-time PCR > Taqman - Human GAPDH Protocol

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

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