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The protocol of the thermal cycler (BIO-RAD, USA) system
| No. of Cycle | Step | Temperature | Time |
|---|---|---|---|
| 1 | Initial denatuartion | 95⁰C | 4 minutes |
| 35 | Denatuartion | 95⁰C | 30 seconds |
| Annealing | 56⁰C | 30 seconds | |
| Extension | 72⁰C | 1 minute | |
| 1 | Final Extension | 72⁰C | 6 minutes |
| - | Hold | 4⁰C | - |
