Objective To establish the methods of EM>in ovo/EM> and EM>ex ovo/EM> culture of chicken embryo as well as EM>in vivo/EM> electroporation. Methods Fertile eggs were incubated for two to three days（E2-E3）, and in ovo electroporation in spinal cord at E2-E3, and ex ovo electroporation in the tectum of brain at E5-E6 for spatial and temporal gene transformation with the parameter such as volt 18V, current 60mA, pause 90ms, and pulse 60ms for six times were carried out. Results The number of samples were 20 eggs for EM>in ovo/EM> culture and 10 eggs for EM>ex ovo/EM> culture. The survival rate of the embryos at E2-E3 was 85% for EM>in ovo/EM> culture and 80% for EM>ex ovo/EM> culture. The number of samples were 11 in spinal cord at E2-E3 ( in ovo electroporation) and 10 in brain tectum at E5-E6 ( ex ovo electroporation). The positive electroporation rate was 54.5% in spinal cord at E2-E3 (EM>in ovo/EM> ) and 60% in brain tectum at E5-E6 (EM>ex ovo/EM>). Conclusion The methods of in ovo and ex ovo culture of chicken embryos and in