Polymorphism of serum transferrin (Tf) in three populations of Huainan spotted yellow chicken was detected by vertical plate polyacryamide gel electrophoresis.Frequencies of genes and genotypes of each population were calculated respectively.Homozygosity of per population and genetic distance among three populations were also analysed The results showed that the frequencies of genes and genotypes of each population were different and the homozygosity of every population should be increased by further selection.