Materials
500 mL:
- 15 g TSB (Bacto)
- 500 ml dH2O
1000 mL:
- 30 g TSB (Bacto)
- 1000 ml dH2O
Procedure
Dissolve medium in water. Add 7.5 g Bacto agar to empty 1 L bottles, aliquot 500 ml of medium to each bottle. Label caps with “TSA” to avoid confusion. Immediately after autoclaving, swirl media to dissolve the melted agar. Tighten caps and place bottles in the water bath. Pour plates when the agar temperature is 50-60 ºC.
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- Add agar directly to the bottles that will be autoclaved, since undissolved agar will be difficult to pour.
- Move bubbles to the side of plates. If there are a lot of bubbles, use a flame to remove them (being careful not to melt the plastic dish).
- It’s important to allow plates enough time to dry out, especially when working with phage. If the underlying agar is too wet, the bacterial lawn might not properly adhere.
- Keep tet plates covered, since tetracycline is light-sensitive.