1. Aspirate medium and wash cell in PBS.
2. Incubate cells at 37oC for 5 min in non-diluted antibiotic-antimycotic.
3. Incubate cells at 37oC for 5 min in trypsin. Spin them down (100g, 5 min). DO NOT EXCEED 100g.
4. Slowly and carefully aspirate supernatant. Resuspend cells in 0.5 ml of regular medium.
NB! We assume that your regular medium contains 1x antibiotic.
5 Transfer cells to a new tube and dilute them with regular medium to 6 ml.
6. Seed cells on 30 mm Petri dishes: 1:1, 1:3, 1:7, 1:15, 1:31, 1:63 and 1:127 (cell suspension/regular medium).
7. Change the medium before you leave for home.
8. Next day: check for results