Streak GFP cells (from -80 freezer) on LB-Kan agar
Grow overnight culture in minimal medium + Kan (50 µg/mL).
(Aliquots in freezer are labeled in 1, 3, 6, 10 mg/mL.)
See here for Kan volumes
Each group needs 4 mL cells. Should prepare to have 8 mL/group = ~100 mL
- Start 3 5 mL overnight cultures the night before.
- Shake at 37C
- Early morning: inoculate culture into minimal medium in flasks, turn off heat.
- Throughout morning: Check OD's.
- If OD > 0.8, dilute
If OD < 0.8, turn on heat
OD should be ~ or just > 0.8 when students come for cells.
Students should get all their solutions ready before they come to get cells.
Dispense cells in glass tubes when students are ready.
For incubation with sugars, use the round bottom centrifuge tubes with the two stage caps from room 368. BD Falcon 352059. 14 mL polypropylene 17 x 100 mm, available at the Fisher stockroom. Fisher 14-959-11B.
During lab, spin the cells down in the big Eppendorf in 368. Don't turn on the refrigeration. Use the 6-tube adapters in the swinging buckets.