Table 1. Thermocycling conditions for Q5 PCR reaction.

Step	Temperature (°C)	Duration	Number of cycles
Initial denaturation	98	30 s	1
Denaturation	98	15 s	30 Back to step 2
Annealing	64	25 s
Extension	72	30 s per kb
Final extension	72	5 min	1
Hold	10	∞	-