Team:Groningen/protocols/Motility assay

From 2013.igem.org

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<h1>Motility assay</h1>
<h1>Motility assay</h1>
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Ad 13 ml of the motility agar (0.4% or 0.7%) per plate.
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Add 13 ml of the motility agar (0.4%) in every petri dish.
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<p>Inoculate the plates at the center with the Strain of interest (strain A).
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<br>Inoculate the plates with the strain of interest, 10 &micro;l with an OD<sub>600</sub> of 0.4.
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<p>For every plate inoculated with Strain A, inoculate a plate with Wild type (WT).
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<br>The plates are inoculated at different temperatures (25&deg;C and 37&deg;C) for 16 hours.
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<p>Inoculations is done with 10 ul of liquid culture with an OD<sub>600</sub> of 0.4.
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<br>After 16 hours of growth, the diameter of the colony is measured.  
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<p>Grow the colonies over 16 hours.
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<p>After the 16 hour growth period the measure the spread of the colony every 2 hours.
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<br><br>
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<p>
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<b>Motility Agar 0.7% (100 ml) </b>
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</p>
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<table id="normal" width="30%">
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  <tr>
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    <th>compound</th>
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    <th>amount</th>
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    <th>treatment</th>
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  </tr>
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  <tr>
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    <td>LB broth </td>
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    <td ALIGN=CENTER>2 g</td>
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    <td></td>
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  </tr>
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  <tr>
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    <td>agar</td>
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    <td ALIGN=CENTER>0.7 g</td>
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    <td></td>
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  </tr>
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</table>
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<br>
 
<p>
<p>
<b>Motility Agar 0.4% (100 ml) </b>
<b>Motility Agar 0.4% (100 ml) </b>
</p>
</p>
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<table id="normal" width="30%">
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<table id="normal" width="60%">
   <tr>
   <tr>
     <th>compound</th>
     <th>compound</th>
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   <tr>
   <tr>
     <td>LB broth </td>
     <td>LB broth </td>
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     <td ALIGN=CENTER>2 g</td>
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     <td>2 g</td>
     <td></td>
     <td></td>
   </tr>
   </tr>
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   <tr>
   <tr>
     <td>agar</td>
     <td>agar</td>
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     <td ALIGN=CENTER>0.4 g</td>
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     <td>0.4 g</td>
     <td></td>
     <td></td>
   </tr>
   </tr>

Revision as of 00:20, 5 October 2013

Motility assay

Add 13 ml of the motility agar (0.4%) in every petri dish.
Inoculate the plates with the strain of interest, 10 µl with an OD600 of 0.4.
The plates are inoculated at different temperatures (25°C and 37°C) for 16 hours.
After 16 hours of growth, the diameter of the colony is measured.

Motility Agar 0.4% (100 ml)

compound amount treatment
LB broth 2 g
agar 0.4 g