Growing cells for 5.2
Growing cells for 5.2 kdorfman Mon, 11/14/2011 - 18:23Cultures for swarm plates
Two days before lab:
Note: C (4130) grows poorly. Start it sooner, in a round bottomed centrifuge tube. Spin down and resuspend so there will be enough cells to inoculate the swarm plate.
Start overnight culture (pick one colony from each Chemotaxis strains , inoculate into ~3 mL Minimal medium + HMLTT.)
Shake at 30C
Day before lab:
First thing in the morning
Check cultures and dilute as necessary. The goal is to have dense cultures by 4 pm.
Late afternoon
Inoculate 52 swarm plates at 4 pm. Incubate at RT. (Students come in at 9 am, take measurements, put plates in 30C incubator, then come back at lab time.)
13 cultures with WT
7 of each of the 6 different Chemotaxis strains.
Start 3 mL overnight cultures of all 7 strains in Minimal medium + HMLTT. Shake at 30C
OR use the same cultures to stab swarm plates as to make usable cultures for motility studies the next day.
Day of lab:
First thing:
Inoculate ~100 µL of each strain into ~25 mL medium (~200 into 50 for wildtype)
During morning and lab:
Check cell concentration, diluting as needed. Do not let cells get to stationary phase!