Decontamination of cells from the yeast

I     Destroy yeast

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.

II     Secure cells from crosscontamination

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.

III    Prepare cell cultures for regular culturing

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