Growing cells for 5.2

Cultures 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!