Sex determination in ratite and non ratite birds by molecular method

In spite of number of methods for sex determination in birds, it is very difficult to distinguish sex especially in ratite birds due to lack of sexual dimorphism. Chromodomain helicase DNA binding 1 gene (CHD 1) is the choice of gene for gender differentiation using PCR based molecular method. In present study, non ratite CHD gene specific primers viz. 1237L/1272H, 2550F/2718R, P2/P8, P2/P3 and ratite bird specific primers viz.W5/ W7 and W1/ K7 were used for gender differentiation in ratite birds. The ratite bird specific primer W5/W7 was the only primer, which determined the sex in emu as well as ostrich successfully, while 1237L/1272H, 2550F/2718R, P2/ P8, P2/P3 primers were unable to discriminate sex in emu and ostrich but ratite and non ratite primers can be used to discriminate the sex in non-ratite bird, primarily in chicken. In an alternative approach of PCR-RFLP, the high resolution melting curve (HRM) analysis showed conflicting pattern in both sexes of ratite birds but in chicken HRM analysis showed clear cut differential melting temperature in both sexes, hence HRM can be used for gender differentiation successfully.