Cell fusion is the central link in hybridoma technology. The basic step is to mix the two cells and add PEG to fuse the cells to each other. The PEG is then diluted with the culture medium to eliminate the effect of PEG. The fused cells were appropriately diluted, and cultured in the wells of the divided culture plates. There are three problems in the fusion process that should be paid special attention. The ratio of 1 cell, the ratio of myeloma cells to spleen cells can vary from 1:2 to 1:10, and the ratio of 1:4 is commonly used. Both cells should be guaranteed to have higher activity prior to fusion. 2 Reaction time, in a mixed cell suspension of the two cells, 4 ml of the culture solution was added dropwise at the first minute; 5 ml of the culture solution was added dropwise at intervals of 2 ml, and then 50 ml of the culture solution was added. 3 The composition of the culture medium, a good culture solution is especially important for the fusion cells, and the calf serum, various ions and nutrients are strictly prepared. If the fusion efficiency is reduced, the media should be checked at any time.

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