Team:Grenoble/Biology/Protocols/LB

From 2012.igem.org

(Difference between revisions)
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 solubilize it</li>
+
         <li>heat the LB Agar up, in order to melt it down</li>
-
         <li>after the solubilization, wait for the solution to refresh</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>
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</section>
</section>
<section>
<section>
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     <h2>Pour the plates</h2>
+
     <h2>Pour into petri dishes</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 into plates:
     <ul>
     <ul>
-
         <li>In a flow hood, pour approximately 15mL of the LB Agar solution into every plates</li>
+
         <li>In a flow hood, pour approximately 15mL of the LB Agar solution into each plate</li>
-
         <li>Let each plate cool until its solid</li>
+
         <li>Let each plate cool until it becomes solid</li>
-
         <li>When it's ok, flip the plates, write on the back and store them at 4°C</li>
+
         <li>When it's ok, flip the plate(bottom-up), write on the back and store them at 4°C</li>
     </ul>
     </ul>
      
      

Revision as of 07:57, 23 August 2012

iGEM Grenoble 2012

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iGEM 2012
main page
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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 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 
 Kanamycin
 Ampicillin

Pour into petri dishes

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 15mL of the LB Agar solution into each plate
  • Let each plate cool until it becomes solid
  • When it's ok, flip the plate(bottom-up), write on the back and store them at 4°C

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