Objective We aimed to investigate the effects of different non-surgical embryo transfer devices, number of transferred embryos, embryo stage, and embryos obtained from different mouse strains on the efficiency of non-surgical embryo transfer in mice, and to compare the efficiencies of surgical and non-surgical embryo transfer, in order to establish a stable non-surgical embryo transfer technology system. Methods Mouse embryo transfer was carried out using non-surgical method . Results The pregnancy rates using two different non-surgical transfer devices were (75.00±0.00)% and (66.67±14.43)%,and the birth rates were (46.11±6.31)% and (18.89±0.96)%,respectively. Transfer of 10, 15, and 20 embryos resulted in pregnancy rates of (66.67±11.55)%, (80.00± 0.00)%, and (66.67±23.09)%, and birth rates of (29.33±4.16)%, (38.67±4.81)%, and (17.00±3.46)%, respectively. When blastocysts and morulae were transferred non-surgically, the resulting pregnancy rates were (80.00±0.00)% and (46.67±11.55)% and the birth rates were (38.67±4.81)% and (10.22±2.77)%, respectively. Four strains (C57BL/6J, ICR, genetically modified mice A, genetically modified mice B) were used as donors for non-surgical embryo transfer, with resulting pregnancy rates of (66.67±11.55)%, (80.00±0.00)%, (73.33±11.55)%, and (80.00±0.00)%, and birth rates of (26.67±2.67)%, (38.67±4.81)%, (32.00±3.53)%, and (29.34±2.31)%, respectively. Fifteen pseudo-pregnant mice were transplanted surgically and 15 were transplanted non-surgically, with pregnancy rates of (80.00±0.00)% and (86.67±11.55)%, and birth rates of (38.67±4.81)% and (36.00±5.82)%, respectively. Conclusions Transfer device A resulted in a higher birth rate in this study. The embryo transfer efficiency was higher when 15 embryos were transferred into unilateral uterine horns of pseudo-pregnant 2.5-day recipients. Blastocyst-stage embryo transfer was more efficient than morula-stage transfer. There was no significant difference in efficiency between surgical and non-surgical embryo transfer procedures.