BACKGROUND:: Whether human leukocyte antigen matching results are accurate or not may directly affect the pairing choice between the donors and recipients. Moreover, the quality of the matching samples may influence the pros and cons of the results. OBJECTIVE:: To establish an experimental technology platform for DNA extraction from the spleen in human leukocyte antigen matching. METHODS:: Lymphocytes were isolated and DNA was extracted from the spleen for the human leukocyte antigen matching, compared with DNA extracted from the whole blood which was normal peripheral blood or hematolytic sample. RESULTS AND CONCLUSION:: Intact electrophoretogram of DNA extracted from the spleen and normal peripheral blood was obtained after amplification and electrophoresis, and the electrophoretogram of DNA extracted from spleen was clearer than that extracted from the normal peripheral blood. However, the intact electrophoretogram of DNA extracted from hematolytic sample was not obtained. An experimental technology platform for DNA extraction from the spleen in human leukocyte antigen matching has been successfully established.