UCSF

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