According to Mendelian heredity laws, the sex ratio of a given chicken population during hatching is expected to be 1:1. In this study, we collected 432 chicken embryos that died during the first week of incubation from 5 different breeds. The sexes of the early-dead embryos were determined by using the previously described molecular sexing technique of double PCR. The female-to-male sex ratio was analyzed for departure from the expected 1:1 sex ratio by chi(2) testing. These results showed that the number of female dead embryos was significantly greater than that of males in the Hubei local breeding stock, Zhusi, and Hy-line Variety Brown (P < 0.05, P < 0.01, P < 0.01 respectively), with observed female-to-male sex ratios of 1.40:1, 2.03:1, and 2.22:1, respectively. Two other Chinese local breeds (the Yellow chicken and the Aijiaohuang chicken) also showed altered sex ratios, although the differences were not significant. Altogether, these results indicated that female chickens were more likely than male chickens to die at the early stages of incubation.