Abstract: The accuracy and the reliability of the association between candidate gene variations and target traits are influenced by the genetic background of papulation, polymorphism of genes, linkage disequilibrium of QTL and the sample size. The association, therefore, should be further confirmed in other populations. However, for the gene with low allelic frequency, it is laborious and cost-consuming to develop a large experimental population to provide adequate individuals with double minor alleles. In the present study, a strategy crossing between heterozygotes was introduced to generate a Mendelian segregating population with larger sample size of the minor homozygous individuals, and accuracy of gene genetic effect evaluation was enhanced due to the closer genetic background and the consideration of family information As an example, in this study, the genetic effect of the pituitary-specific transcription factor gene (POU1F1-6) on chicken growth rate was confirmed by this strategy.The results indicated that the strategy was applicable, laborsaving, and cost-effective.