1. 75 seconds at 95˚C

2. a―45 seconds at 94˚C

b―45 seconds at 62˚C

c―60 seconds at 72˚C

repeat step 2. for 4 cycles

3. a―45 seconds at 94˚C

b―45 seconds at 61˚C

c―60 seconds at 72˚C

repeat step 3. for 4 cycles

4. a―45 seconds at 94˚C

b―45 seconds at 60˚C

c―60 seconds at 72˚C

repeat step 4. for 4 cycles

5. a―45 seconds at 94˚C

b―45 seconds at 59˚C

c―60 seconds at 72˚C

repeat step 5. for 4 cycles

6. a―45 seconds at 94˚C

b―45 seconds at 58˚C

c―60 seconds at 72˚C

repeat step 6. for 4 cycles

7. a―45 seconds at 94˚C

b―45 seconds at 57˚C

c―60 seconds at 72˚C

repeat step 7. for 4 cycles

8. a―45 seconds at 94˚C

b―45 seconds at 56˚C

c―60 seconds at 72˚C

repeat step 8. for 4 cycles

9. a―45 seconds at 94˚C

b―45 seconds at 55˚C

c―60 seconds at 72˚C

repeat step 9. for 4 cycles

10. a―45 seconds at 94˚C

b―45 seconds at 54˚C

c―60 seconds at 72˚C

repeat step 10. for 4 cycles

11. a―45 seconds at 94˚C

b―45 seconds at 53˚C

c―60 seconds at 72˚C

repeat step 11. for 4 cycles

12. a―45 seconds at 94˚C

b―45 seconds at 52˚C

c―60 seconds at 72˚C

repeat step 12. for 20 cycles