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 of overnight culture 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 spot is dried for half an hour, after which 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>Let the colonies grow 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>
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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>
     <th>amount</th>
     <th>amount</th>
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    <th>treatment</th>
 
   </tr>
   </tr>
   <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>
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    <td></td>
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   </tr>
   </tr>
   <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>
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    <td></td>
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   </tr>
   </tr>
</table>
</table>

Latest revision as of 00:22, 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 of overnight culture with an OD600 of 0.4).
The spot is dried for half an hour, after which 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
LB broth 2 g
agar 0.4 g