Harvest Yeast Cells
- Grow cell culture to late-log overnight in 5mL of YEPD.
- Subclone 1:100 into a flask containing the desired amount of media (50 mL - 2L).
- Grow until mid-log phase.
- Transfer media to a 50mL conical vial for smaller cultures (100mL or less) or into a 500mL centrifuge bottle for larger cultures.
- Spin down at 4k RPM and 4°C for 3-5 minutes.
- Discard supernatant and refill with any remaining media. Spin down again and continue until all of the cell culture has been pelleted.
- Use 1x volume of PBS for TAP purification or CoIP.
- Use 1x volume of TBS for ChIP.
- Use 1x volume of PBS for RNA extraction. Note that, for RNA extraction, all steps for harvest must be performed at room temperature, including any centrifugation.
- Discard the wash supernatant and resuspend the cell pellet in any remaining PBS. It may be necessary to add a small amount of additional PBS.
- Transfer the resuspended cell pellet into a 2 mL FastPrep screw-top tube, or into a 50 mL conical vial. Do not overtighten caps.
- Spin down at 4k RPM and 4°C for 3-5 minutes for 50 mL conicals or flash spin for 2 mL FastPrep tubes. Discard any remaining supernatant.
- Immerse in liquid nitrogen until frozen.
- Transfer to -80°C Freezer.