User:Kalkao/notebook/Streaking Plates

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  1. Obtain cell stocks from the -80*C storage and place in a cold block
  2. Obtain T-broth agar plates from the 4*C fridge
  3. Use a sterile loop to streak cells onto the plates
    • Use fireboy to flame sterile loop
    • Dip the loop approximately 1/4 inch into the BL21 stock
    • Spread three lines of stock culture onto one side of the agar plate
    • Spread three lines of the culture on the plate across the top of the agar with the sterile loop
    • Rotate the plate 90* and spread three lines of culture across the top of the agar again from the existing streaks
    • Rotate the plate 90* and make three final streaks across the top of the agar
    • Be sure to flame the sterile loop between each set of three streaks
    • After flaming the sterile loop, cool the loop by touching it to the agar before you streak cells
    • Remember to use good sterile technique so that the plates do not become contaminated
    • Repeat this step for each stock on a new T-broth agar plate
  4. When streaking is finished, put the plates in the 30*C incubation room to allow the colonies to form slowly
    • The colonies should be ready in approximately 15 hours