Relatively new to splitting cells in tissue culture. I am splitting cells on Monday and Fridays, with a 1:10 split.
My current advised protocol (just for amounts) is as follows:
1) Remove spent media from T25 flask containing cells
2) Add 5-10ml PBS, swirl to wash
3) Remove all PBS
4) Add 2ml TrypLe and ensure complete coverage
5) Incubate for 2-5 minutes
6) Remove T25 flask from incubator and take 200ul from old T25 flask and put into new T25 flask
7) Add 5ml media
8) Incubate until next split
I just wanted to make sure this protocol is correct in terms of the amounts, I have done this a couple times so far and cells seem to still be alive etc. The concept isn't complicated however I seem to find it difficult to get my head around sometimes.
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