Table 4. PCRs reactions thermal profiles.

Step	ESR1 – first amplicon		ESR1 – second amplicon
	duration	cycles no.	duration	cycles no.
Initial denaturation	95°C, 5 min	1	95°C, 5 min	1
Denaturation	95°C, 20 sec	30	95°C, 30 sec	40
Annealing	71°C, 15 sec	30	60°C, 30 sec	40
Elongation	72°C, 15 sec	30	70°C, 1.20 sec	40
Final elongation	72°C, 3 min	1	70°C, 5 min	1