RT-PCR
|
Sample |
RNA amount add (1ug) |
DEPC water (ul) |
Primer (Oligo(dT)) (ul) |
10mM dNTP mix (ul) |
Total (ul) |
|||
|
A |
x ul |
9-x ul |
1 |
2 |
12 |
|||
65°C for 5 min in PCR machine
4°C in PCR machine
Master Mix
|
1X |
20X |
|||||||
|
5x cDNA Synthesis buffer (vortex) |
4ul |
80ul |
||||||
|
0.1M DTT |
1ul |
20ul |
||||||
|
RNAse OUT |
1ul |
20ul |
||||||
|
DEPC-H2O |
1ul |
20ul |
||||||
|
ThermoScript RT |
1ul |
20ul |
||||||
|
Total |
8ul |
160ul |
||||||
Add 8ul of the Master Mix to each sample
50°C for 30 min and 85°C for 5 min in PCR machine.
Store at -20°C