Abstract: In order to calibrate the gamma spectrum response curves for gamma and neutron detection, a series of quasi-single energy gamma radioactive sources with energy range from 100 to 2 700 keV must be used in the experiments. The paper mainly concerns in the method of preparation of quasi-single energy gamma radioactive serial source by using the fast rabbit irradiation system on Xi’an pulse reactor. The influencing factors in irradiation process were discussed, and the solutions for reducing influencing factors were separately given for different irradiated samples. The measuring means of activity were introduced for irradiated samples with high and low radioactive source respectively, and the self-absorption coefficients of all measured samples were calculated.