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Team:Grenoble/Biology/Protocols/LB
From 2012.igem.org
(Difference between revisions)
| Line 48: | Line 48: | ||
<section> | <section> | ||
<h2>Pour the plates</h2> | <h2>Pour the plates</h2> | ||
| - | When the antibiotic solution is added or when the bottle is not so hot (around 50°C), you can 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: |
| - | + | <ul> | |
| - | + | <li>In a flow hood, pour approximately 15mL of the LB Agar solution into every plates</li> | |
| + | <li>Let each plate cool until its solid</li> | ||
| + | <li>When it's ok, flip the plates, write on the back and store them at 4°C</li> | ||
| + | </ul> | ||
</section> | </section> | ||
Revision as of 07:45, 23 August 2012
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 sterelize 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 added 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
