Versene with trypsin

Very often, however, mixtures of trypsin or trypsin-citrate with versene are used to release cells from monolayer cultures as the combination works very much faster than either alone. In our laboratory the following procedure is used to make up 10 1 of versene solution in PBS-A ... 

Treat the fixed preparation with trypsin (0.025%) versene pH7.0 for 10-15 min at 30°C. This causes considerable swelling of the chromosomes. The actual time of treatment should be monitored by phase contrast microscopy — the end point being when the two chromatids fuse together and appear as a unit. 

F9 and similar EC cells are sensitive to trypsinisation and this sensitivity can be reduced by treating with a 0.025% trypsin versene... 

Remove media and trypsinize cells with 0.05% trypsin versene (1 1) until all cells have rounded up and detached (see Note 4). 

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