User:Kalkao/notebook/Making Overnight Suspensions

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  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
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