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Team:Grenoble/Biology/Protocols/LB
From 2012.igem.org
LB Agar plates preparation
Goal
Prepare solid medium to culture bacteria.Prepare the LB Agar
In a 500mL bottle add 6.4g of bacteriological agar to 400mL of LB medium. Then sterilize it using the autoclave.You can store it at room temperature, but before use it, you need to heat it up.
Add the right antibiotic
If you want to prepare LB Agar plates with antibiotics you need to:- Heat the LB Agar up, in order to melt it down.
- After the melting, wait for the solution to cool down.
- When the solution is not so hot (around 50°C), add the antibiotic solution (under a flow hood of course).
Antibiotics concentrations:
| Antibiotics | Finale concentration |
|---|---|
| Chloramphenicol | 0.03 10-3 g/mL |
| Kanamycin | 0.05 10-3 g/mL |
| Ampicillin | 0.1 10-3 g/mL |
Pour into plates
When the antibiotic solution is added or when the bottle is not so hot (around 50°C), you can pour into plates:- In a flow hood, pour approximately 15 mL of the LB Agar solution into each plate.
- Let each plate cool until it becomes solid.
- When it's ok, flip the plates (bottom-up), write on the back and store them at 4°C.
