Abstract: We studied the polyacrylamide gel-casting new and old glass plates and gasket in the preparation of small amount of separation gel to seal on polyacrylamide gel plate (Do it yourself, DIY) and compared the effects thirty times each of them. After that the DIY gel was used to electrophorese the protein samples of Jurkat and A549 cell lines, then strain the transformed PVDF with Coomassie blue dye and detect Actin antibody reaction with Western blot. The results showed that the success rate of gel-casting new plate and gasket was the best to whole or a part of the old those with statistically significant difference (P <0.05). But there was no statistical difference between the old method and that of the DIY(P >0.05). As same time the DIY gel was used in the subsequent experiments. It was normal that the protein samples in electrophoresis gel and PVDF Coomassie blue staining. The integrated intensities of Actin expression of two cell line were the same (P >0.05). So by the DIY method could prolong the service life of gel-casting plate and gasket with funds retrenched. It had become a simple and practical method to prepare polyacrylamide gel plate.