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Resources

Mitomycin treatment of PMEF cells

1
Disolve 1 bottle of Mitomycin C (Sigma, cat#……) in 200ml EF medium (keep lights off under the hood)
2
Add 20ml/flask (use 150cm2 flasks) of EF+mitomycin medium
3
Incubate 2h at 37C
4
Wash with PBS twice
5
Add 5ml trypsin/flasks
6
Incubate at RT for 5 minutes
7
Resuspend in 10 ml of EF medium
8
Spin 5 min
9
Freez in 2 tubes, use 1ml of freezing medium per tube