How do you make ampicillin plates?
Simply measure out 100 mg of ampicillin powder, add it to 1 mL of water, dissolve by vortexing, and filter sterilize. Prepare a water bath at 60 ℃ with sufficient water to submerge ~75% of the bottle containing your molten gel mixture.
How much agar do you use for plates?
Use about 30 mL of the agar-medium mix to create each plate when using 100-mm diameter dishes . The less agar-medium mix in each plate, the more easily they will dry out. 30 mL is a good amount for long-term storage, 10–20 mL is fine if you are going to use the plates relatively soon.
How do you make LB agar ampicillin plates?
To make 500 ml of LB agar containing 50 µg/ml ampicillin, add 2.5 ml of 10 mg/ml ampicillin stock. Ensure LB agar is cooled to 50°C before adding ampicillin, excessive heat will degrade the ampicillin. Swirl or use the stir bar and a stir plate to mix the ampicillin into the agar taking care not to introduce bubbles.
How much agar do I need for 20 plates?
This recipe is for 500 mL of LB agar. This makes about 20 plates (1 bag).
How do you get rid of condensation on agar plates?
A significant reduction in the amount of condensation can be achieved by allowing the sterilized agar to cool to 50°C–55°C before pouring. A water bath set at 50°C is useful to store bottles of molten agar to maintain the optimal temperature prior to pouring plates. Remember agar solidifies at around 42°C.
How do you make LB agar plates from LB broth?
Protocol
- 500 mL makes about one sleeve of petri dishes.
- Add 25g LB Broth per Liter to an appropriately sized beaker.
- Add Water and Stir until clumps are gone.
- Add 15 g Agar per Liter , this will not dissolve.
- Transfer to a erlenmeyer flask at least 2x the volume that you are making.
How do you make 50mg mL ampicillin?
A 50 mg/ml stock solution of ampicillin can be prepared by dissolving 500 mg ampicillin sodium salt in water to a final volume of 10 ml.