Table 2.

PCR mixture content and thermal profile of the reactions.

PCR reaction mixture
Component	Initial concentration	Volume added	Final concentration	Mixture volume
ZymoTaq™ Premix	2x	5 µl	1x	10 µl
→PCR	10 µM	1 µl	1 µM
←PCR	10 µM	1 µl	1 µM
DNA	100 ng/µl	0,2 µl	2 ng/µl
nuclease-free water		2,8 µl
Thermal profile of the reactions
Number of cycles	Step		Duration, temperature
1	initial denaturation		10 min., 95 °C
37	denaturation		30 s., 95 °C
	annealing		30 s., 54 °C
	extension		60 s., 72 °C
1	final extension		7 min., 72 °C
1	hold		∞, 4 °C

→PCR, forward primer; ←PCR, reverse primer; min., minutes, s., seconds.