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Team:Grenoble/Biology/Protocols/LB
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<h2>Prepare the LB Agar</h2> | <h2>Prepare the LB Agar</h2> | ||
In a 500mL bottle add 6.4g of bacteriological agar to 400mL of LB medium. | In a 500mL bottle add 6.4g of bacteriological agar to 400mL of LB medium. | ||
| - | Then | + | Then sterilize it using the autoclave.<br/> |
You can store it at room temperature, but before use it, you need to heat it up. | You can store it at room temperature, but before use it, you need to heat it up. | ||
</section> | </section> | ||
<section> | <section> | ||
<h2>Add the right antibiotic</h2> | <h2>Add the right antibiotic</h2> | ||
| - | If you want to prepare LB Agar plates | + | If you want to prepare LB Agar plates with antibiotics you need to: |
<ul> | <ul> | ||
| - | <li> | + | <li>Heat the LB Agar up, in order to melt it down.</li> |
| - | <li> | + | <li>After the melting, wait for the solution to cool down.</li> |
| - | <li> | + | <li>When the solution is not so hot (around 50°C), add the antibiotic solution (under a flow hood of course).</li> |
</ul> | </ul> | ||
<br/> | <br/> | ||
| Line 33: | Line 33: | ||
<tr> | <tr> | ||
<td> Chloramphenicol </td> | <td> Chloramphenicol </td> | ||
| - | <td></td> | + | <td> 0.03 10<span class="exposant">-3</span> g/mL</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td> Kanamycin</td> | <td> Kanamycin</td> | ||
| - | <td></td> | + | <td> 0.05 10<span class="exposant">-3</span> g/mL</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td> Ampicillin</td> | <td> Ampicillin</td> | ||
| - | <td></td> | + | <td> 0.1 10<span class="exposant">-3</span> g/mL</td> |
</tr> | </tr> | ||
</table> | </table> | ||
| Line 47: | Line 47: | ||
</section> | </section> | ||
<section> | <section> | ||
| - | <h2>Pour | + | <h2>Pour into plates</h2> |
| - | When the antibiotic solution is added or when the bottle is not so hot (around 50°C), you can pour | + | When the antibiotic solution is added or when the bottle is not so hot (around 50°C), you can pour into plates: |
| - | + | <ul> | |
| - | + | <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>When it's ok, flip the plates (bottom-up), write on the back and store them at 4°C.</li> | ||
| + | </ul> | ||
</section> | </section> | ||
Latest revision as of 16:35, 20 September 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 (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.
