Strategy for embryo sex identification in the yak (Bos grunniens) by simultaneous amplification of the SRY and bHBB genes

Figures & data

Table 1. Primer sequences and PCR product size for sex identification in yak.

Gene	Upstream primers	Downstream primers	Size of product (bp)
bHBB inner	5′-CATCCTCAGAGCCCAAAG-3′	5′-CGACTAGCCCTTCATTTCT-3′	184
Outer	5′-GACTTTCAGAAGGTGGTG-3′	5′-GAGTAAGGCTAAGGAGCA-3′	373
SRY inner	5′-ACGAAGACGAAAGGTG-3′	5′-TGTATGTGAAGGGGTG-3′	272
Outer	5′-AACGCCTTCATTGTGT-3′	5′-CTGCTGTGATGCTCCT-3′	404

Figure 1.  Identification of four pair primers. M: DL2000 DNA marker. Lanes 1–4 contain male yak DNA and outer primer (404 bp) or inner primer (272 bp) of SRY gene. Lanes 5–8 contain female yak DNA and outer primer (373 bp) or inner primer (184 bp) of bHBB gene.

Figure 2.  Optimisation of PCR protocol for sex identification in the yak by simultaneous amplification of the SRY (272 bp) and bHBB (184 bp) genes. An aliguot (1 µL) of the first PCR product was submitted to 30 cycles in the nest amplification containing 1.5 µL each inner primer (0.6 µM) of SRY and bHBB. (a) The nested PCR products amplified with various concentrations of male yak DNA and outer primer in the first PCR. 1–3: DNA 15, 10, and 5 ng/µL; 4–7: outer primer 0.2, 0.4, 0.6 and 0.8 µM; M: DL2000 marker. (b) The nested PCR products amplified with various cycles of the first PCR. 1–4: 15 cycles; 5–8: 20 cycles; 9–12: 25 cycles; 13–16: 30 cycles; M: DL2000 DNA marker. Lanes 1, 2, 5, 6, 9, 10, 13, 14 contain male yak DNA; Lanes 3, 4, 7, 8, 11, 12, 15, 16 contain female yak DNA.