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Team:Grenoble/Biology/Protocols/LB
From 2012.igem.org
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| Line 20: | Line 20: | ||
If you want to prepare LB Agar plates with antibiotics you need to: | If you want to prepare LB Agar plates with antibiotics you need to: | ||
<ul> | <ul> | ||
| - | <li>Heat the LB Agar up, in order to melt it down</li> | + | <li>Heat the LB Agar up, in order to melt it down.</li> |
| - | <li>After the melting, wait for the solution to cool down</li> | + | <li>After the melting, wait for the solution to cool down.</li> |
| - | <li>When the solution is not so hot (around 50°C), add the antibiotic solution (in a flow hood of course)</li> | + | <li>When the solution is not so hot (around 50°C), add the antibiotic solution (in a flow hood of course).</li> |
</ul> | </ul> | ||
<br/> | <br/> | ||
| Line 50: | Line 50: | ||
When the antibiotic solution is added or when the bottle is not so hot (around 50°C), you can 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: | ||
<ul> | <ul> | ||
| - | <li>In a flow hood, pour approximately | + | <li>In a flow hood, pour approximately 15 mL of the LB Agar solution into each plate.</li> |
<li>Let each plate cool until it becomes solid.</li> | <li>Let each plate cool until it becomes solid.</li> | ||
<li>When it's ok, flip the plates (bottom-up), write on the back and store them at 4°C.</li> | <li>When it's ok, flip the plates (bottom-up), write on the back and store them at 4°C.</li> | ||
Revision as of 08:48, 28 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 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 (in 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.
