PCR Amplification (E6120)
Protocol
- Mix the following components in a sterile PCR tube:
RT reaction mixture: 20 μl
LongAmp Taq® 2X Master Mix: 25 μl
SR Primer F1: 2.5 μl
SR Primer R1: 2.5 μl
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Total volume: 50 μl - PCR cycling conditions:.
Cycle step Temp Time Cycles Initial denaturation 94°C 30 sec 1 Denaturation
Annealing
Extension94°C
60°C
65°C10 sec
30 sec
15 sec12* Final extension 65°C 5 min 1 Hold 4°C