User:Kalkao/notebook/Making Overnight Suspensions

1. Retrieve streaked plates the 30*C incubation room
   • If colonies need more time to form, place in the 37*C incubation room for faster growth
   • Plates with colonies can be kept in the 4*C fridge until for awhile
2. Place your plates on the lab bench
3. Obtain one test tube for each plate streaked
4. Sterilize a pipette:
   • Using a fireboy, flame the top of the pipette
   • Place the pipet into the pipetteman
   • Now, slowly flame the entire length of the pipette
5. Pipette 5mL of T-broth into a test tube
   • Make sure the cap is on the T-broth bottle when it is not in use
   • Flame the opening of the T-broth bottle directly before and after the pipette enters the bottle
   • Flame the opening of the test tube before and after the T-broth is added
   • Dispose of the pipette in the bleach water
6. Use the sterile loop to move a single colony from one plate to one test tube with T-broth
   • Carefully touch one colony with the sterile loop
   • Put sterile loop into the T-broth in the test tube
   • Use the loop to mix the T-broth
   • Vortex the test tube
   • Flame the sterile loop before it touches the agar plate and after it leaves the test tube
   • Remember to flame the entire length of the sterile loop for this step because the much of the wire will be inside the test tube when mixing the T-broth
   • Flame the opening of the test tube directly before and after the sterile loop is inside
   • Repeat this step for every streaked plate
7. Bring the test tube cultures to the 37*C incubation room
8. Stop the rotating tube rack
9. Place test tube with cell cultures on the tube rack
   • Make sure the rack is balanced
10. Turn the rotor back on and set the dial between 7 and 8
11. Parafilm the streaked plates and place in the 4*C fridge