Frequency of heterozygote individuals (n = 122), separated by sex (63 males: light blue, 59 females: pink) for markers selected by their segregation pattern in the larger mapping family and by their position on Pun-LG10 (Ore-LG23). A) 78 markers, selected as reciprocally homozygous (AA/BB) in the F0 and heterozygote in both, the male and the female F1 (AB/AB), segregate as expected in a 50:50 ratio of AA:AB in F2 females and AB:BB in F2 males, resulting in frequency of heterozygous F2 individuals around 0.5 for both sexes. B) 70 markers, selected as homozygote in the F0 and F1 females and heterozygote in the F0 and F1 males, segregate similarly in the F2; i.e., the frequency of heterozygous individuals is approaching 0 in females and 1 in males for positions < 35 Mb. The QTL region (Bayesian confidence interval) for sex determination in Pundamilia is located between 27.8 and 29.7 Mb (shaded region).