Hemoglobin D-Punjab is a common Hb variant in China. This paper describes a new way——Eco RI mapping of the amplified β-globin DNA, for identification of Hb D-Punjab gene. The primers for PCR were designed and synthesized to enzymatically amplify an 144bp fragment which contained an Eco R1 recognition site. So the D-Punjab gene could be easily detected by Eco RI digestion of the amplified sequences on agarose gel electrophoresis owing to a single base change on codon 121. Four Hb DPunjab families from Han, Tibet and Kasak nationalities were analysed by this simple method. This results were also confirmed by oligonucleotide hybridization technique.