Table 3.

Optimized amplification conditions for multiplex PCR for RYSC3, M6 and M45 makers

	Final concentration	PCR conditions
NFW		Start: 94 °C × 1 min
M6F1	0.6 µM	30 cycles of 3 steps: 94 °C × 30 s
M6R4	0.6 µM	60 °C × 1 min
ADG23R	0.1 µM	65 °C × 4 min
3.3.3 s	0.1 µM	Last:
M45F1	0.3 µM	65 × 60 s
M45R1	0.3 µM
10X PCR buffer (25 mM MgCl2)	1×
dNTPs (5 mM)	0.2 mM
Taq polymerase	0.5 U
Genomic DNA	10–100 ng