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Team:Grenoble/Biology/Protocols/LB
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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 solubilize it
- after the solubilization, wait for the solution to refresh
- when the solution is not so hot (around 50°C), add the antibiotic solution (in a flow hood of course)
Antibiotics concentrations:
| Antibiotics | Finale concentration |
|---|---|
| Chloramphenicol | |
| Kanamycin | |
| Ampicillin |
Pour the plates
When the antibiotic solution is added or when the bottle is not so hot (around 50°C), you can pour the plates:- In a flow hood, pour approximately 15mL of the LB Agar solution into every plates
- Let each plate cool until its solid
- When it's ok, flip the plates, write on the back and store them at 4°C
