Thereafter, 11 polymorphic SSR
and InDel markers (Table 1) located in the critical
genomic region were identified, and these, along with
the most closely linked SSRs identified in the primary
screen, were used to genotype a population of 1 720
F2 seedlings to obtain a localized fine-scale genetic
map. Each 10 μL PCR contained 1 μL of template DNA,
1 μL of 10 × PCR buffer, 0.1 μL of dNTPs (10 μmol/L),
1 μL of primers (10 μmol/L) and 0.1 U Taq DNA
polymerase. The amplification protocol comprised an
initial denaturation (95 °C for 3 min), followed by 35
cycles of 95 °C for 30 s, 55 °C for 30 s and 72 °C for