User:Kalkao/notebook/Streaking Plates
From OpenWetWare
< User:Kalkao | notebook
- Obtain cell stocks from the -80*C storage and place in a cold block
- Obtain T-broth agar plates from the 4*C fridge
- 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
- 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