The monomorphism of newly hatched chicks poses a problem for farmers who need to separate them. Currently, the most widely used technique is the cloacal sexing method which has a low accuracy rate. Thus, a PCR based gender determination protocol was established to obtain a higher accuracy rate than conventional sexing methods. Feathers of a day up to a week old chicks were collected for the molecular analysis using the PCR targeting the CHD-W and CHD-Z genes in the chicken's chromosome Z and chromosome W, respectively. The results of this study showed that the PCR based gender determination protocol was a sensitive and accurate method for determining the gender of monomorphic chicks as compared to the conventional sexing method.