RT-PCR Using NOVASCRIPT-III-SS Kit
Reaction Mixture:
2X One-Step RT-PCR Buffer 25.0µl
One Step Enzyme Mix 2.0µl
Forward Primer (50µl) 0.2µl
Reverse Primer (50µl) 0.2µl
RNA Sample (500ng) 1.0µl
RNase Inhibitor(10 Units) 1.0µl
DEPC-Treated Water 20.6µl
PCR-Program:
1. T= 42 30min
2. T= 95 10min
3. T= 95 30sec
4. T= 55 30sec
5. T= 72 1:30min (Depending on Resulting Amplicon Size, 1min for 1kb DNA)
6. GO TO 3 REP 35
7. T= 72 7min
8. HOLD 4