Team:Groningen/protocols/biofilm media

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<div class="mainContent">
<div class="mainContent">
-
<h2>Media for macrocolony formation</h2>
+
Make an overnight culture of the strains of interest in LB broth.
 +
<br>1 ml of the O/N culture is required for every 10 ml of Sgg medium (2x SG supplemented with 1% [wt/vol] glycerol).
 +
<br>This will be incubated at 27&deg;C for 5 days.
-
<br><H5>2x SG medium (100mL)</H5>
+
<h5>Media for macrocolony formation</h5>
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<table border="1">
+
<H5 id="special">2x SG Medium (100 ml)</H5>
 +
<table id="normal" width="70%">
   <tr>
   <tr>
     <th>compound</th>
     <th>compound</th>
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   <tr>
   <tr>
     <td>Nutrient broth (Difco)</td>
     <td>Nutrient broth (Difco)</td>
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     <td ALIGN=CENTER>1.6g</td>
+
     <td>1.6 g</td>
     <td></td>
     <td></td>
   </tr>
   </tr>
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   <tr>
   <tr>
     <td>KCl</td>
     <td>KCl</td>
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     <td ALIGN=CENTER>0.2g</td>
+
     <td>0.2 g</td>
     <td></td>
     <td></td>
   </tr>
   </tr>
   <tr>
   <tr>
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     <td>MgSO<sub>4</sub>7H<sub>2</sub>O </td>
+
     <td>MgSO<sub>4</sub>.7H<sub>2</sub>O </td>
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     <td ALIGN=CENTER>0.05g</td>
+
     <td>0.05 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>1.5g</td>
+
     <td>1.5 g</td>
-
     <td> </td>
+
     <td></td>
   </tr>
   </tr>
<tr>
<tr>
     <td></td>
     <td></td>
-
     <td ALIGN=CENTER></td>
+
     <td></td>
-
     <td> add al together and autoclave</td>
+
     <td> add everything together and autoclave</td>
   </tr>
   </tr>
<tr>
<tr>
-
     <td>Ca(NO<sub>3</sub>)<sub>2</sub>4H<sub>2</sub>O  1mM</td>
+
     <td>Ca(NO<sub>3</sub>)<sub>2</sub>.4H<sub>2</sub>O  (1 mM)</td>
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     <td ALIGN=CENTER>0.1mL</td>
+
     <td>0.1 ml</td>
-
     <td> </td>
+
     <td></td>
   </tr>
   </tr>
<tr>
<tr>
-
     <td>MnCl<sub>2</sub>4H<sub>2</sub>O  0.1mM</td>
+
     <td>MnCl<sub>2</sub>.4H<sub>2</sub>O  (0.1 mM)</td>
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     <td ALIGN=CENTER>0.1mL</td>
+
     <td>0.1 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
<tr>
<tr>
-
     <td>FeSO<sub>4</sub>  1 uM</td>
+
     <td>FeSO<sub>4</sub>  (1 &micro;M)</td>
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     <td ALIGN=CENTER>0.1mL</td>
+
     <td>0.1 ml</td>
-
     <td> </td>
+
     <td></td>
   </tr>
   </tr>
<tr>
<tr>
     <td>0.1% glucose </td>
     <td>0.1% glucose </td>
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     <td ALIGN=CENTER>0.5mL</td>
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     <td>0.5 ml</td>
-
     <td> </td>
+
     <td></td>
   </tr>
   </tr>
</table>  
</table>  
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<br><H5>LB/Mn/glucose medium (100mL)</H5>
+
<br><H5 id="special">LB/Mn/Glucose Medium (100 ml)</H5>
-
<table border="1">
+
<table id="normal" width="70%">
   <tr>
   <tr>
     <th>compound</th>
     <th>compound</th>
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   <tr>
   <tr>
-
     <td>LB/ 1.5% agar</td>
+
     <td>LB 1.5% agar</td>
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     <td ALIGN=CENTER>100ml</td>
+
     <td>100 ml</td>
     <td>autoclave</td>
     <td>autoclave</td>
   </tr>
   </tr>
<tr>
<tr>
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     <td>MnCl<sub>2</sub>4H<sub>2</sub>O  0.1mM</td>
+
     <td>MnCl<sub>2</sub>.4H<sub>2</sub>O  (0.1 mM)</td>
-
     <td ALIGN=CENTER>0.1mL</td>
+
     <td>0.1 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
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<tr>
<tr>
     <td>0.1% glucose </td>
     <td>0.1% glucose </td>
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     <td ALIGN=CENTER>0.5mL</td>
+
     <td>0.5 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
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<br><H5> MSgg medium (100mL)</H5>
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<br><H5 id="special"> MSgg Medium (100 ml)</H5>
-
<table border="1">
+
<table id="normal" width="70%">
   <tr>
   <tr>
     <th>compound</th>
     <th>compound</th>
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   <tr>
   <tr>
     <td>KH<sub>2</sub>PO<sub>4</sub></td>
     <td>KH<sub>2</sub>PO<sub>4</sub></td>
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     <td ALIGN=CENTER>0.026g</td>
+
     <td>0.026 g</td>
     <td></td>
     <td></td>
   </tr>
   </tr>
<tr>
<tr>
     <td>K<sub>2</sub>HPO<sub>4</sub></td>
     <td>K<sub>2</sub>HPO<sub>4</sub></td>
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     <td ALIGN=CENTER>0.061g</td>
+
     <td>0.061 g</td>
     <td></td>
     <td></td>
   </tr>
   </tr>
   <tr>
   <tr>
-
     <td>MOPS 100mM</td>
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     <td>MOPS (100 mM)</td>
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     <td ALIGN=CENTER>2.09g</td>
+
     <td>2.09 g</td>
     <td></td>
     <td></td>
   </tr>
   </tr>
   <tr>
   <tr>
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     <td>MgCl<sub>2</sub>6H<sub>2</sub>O  2mM</td>
+
     <td>MgCl<sub>2</sub>.6H<sub>2</sub>O  (2 mM)</td>
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     <td ALIGN=CENTER>0.04g</td>
+
     <td>0.04 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>1.5g</td>
+
     <td>1.5 g</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
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<tr>
<tr>
     <td></td>
     <td></td>
-
     <td ALIGN=CENTER></td>
+
     <td></td>
-
     <td> add al together and autoclave</td>
+
     <td>add everything together and autoclave</td>
   </tr>
   </tr>
<tr>
<tr>
-
     <td>CaCl<sub>2</sub>  700mM</td>
+
     <td>CaCl<sub>2</sub>  (700 mM)</td>
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     <td ALIGN=CENTER>0.1mL</td>
+
     <td>0.1 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
<tr>
<tr>
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     <td>MnCl<sub>2</sub> 50uM</td>
+
     <td>MnCl<sub>2</sub> (50 &micro;M)</td>
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     <td ALIGN=CENTER>50uL</td>
+
     <td>50 &micro;l</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
<tr>
<tr>
-
     <td>FeCl<sub>3</sub>  50 uM</td>
+
     <td>FeCl<sub>3</sub>  (50 &micro;M)</td>
-
     <td ALIGN=CENTER>0.1mL</td>
+
     <td>0.1 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
<tr>
<tr>
-
     <td>ZnCl<sub>2</sub> 1 uM</td>
+
     <td>ZnCl<sub>2</sub> (1 &micro;M)</td>
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     <td ALIGN=CENTER>0.1mL</td>
+
     <td>0.1 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
<tr>
<tr>
-
     <td>thiamine 2 uM</td>
+
     <td>thiamine (2 &micro;M)</td>
-
     <td ALIGN=CENTER>0.1mL</td>
+
     <td>0.1 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
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<tr>
<tr>
     <td>0.5% glycerol </td>
     <td>0.5% glycerol </td>
-
     <td ALIGN=CENTER>0.57mL</td>
+
     <td>0.57 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
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<tr>
<tr>
     <td>0.5% glutamate </td>
     <td>0.5% glutamate </td>
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     <td ALIGN=CENTER>10mL</td>
+
     <td>10 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
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<tr>
<tr>
     <td>tryptophan </td>
     <td>tryptophan </td>
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     <td ALIGN=CENTER>1mL</td>
+
     <td>1 ml</td>
     <td> </td>
     <td> </td>
   </tr>
   </tr>
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</p>
</p>
-
<br>Grow cells overnight in LB / glass tubes at 37C.
 
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<br>Inoculate 2uL spot on plate dry 15 minutes at 30C
 
-
<br>Inoculate 30C 2-4 days
 

Latest revision as of 00:27, 5 October 2013
Make an overnight culture of the strains of interest in LB broth.
1 ml of the O/N culture is required for every 10 ml of Sgg medium (2x SG supplemented with 1% [wt/vol] glycerol).
This will be incubated at 27°C for 5 days.
Media for macrocolony formation
2x SG Medium (100 ml)
compound amount treatment
Nutrient broth (Difco) 1.6 g
KCl 0.2 g
MgSO4.7H2O 0.05 g
agar 1.5 g
add everything together and autoclave
Ca(NO3)2.4H2O (1 mM) 0.1 ml
MnCl2.4H2O (0.1 mM) 0.1 ml
FeSO4 (1 µM) 0.1 ml
0.1% glucose 0.5 ml

LB/Mn/Glucose Medium (100 ml)
compound amount treatment
LB 1.5% agar 100 ml autoclave
MnCl2.4H2O (0.1 mM) 0.1 ml
0.1% glucose 0.5 ml

MSgg Medium (100 ml)
compound amount treatment
KH2PO4 0.026 g
K2HPO4 0.061 g
MOPS (100 mM) 2.09 g
MgCl2.6H2O (2 mM) 0.04 g
agar 1.5 g
add everything together and autoclave
CaCl2 (700 mM) 0.1 ml
MnCl2 (50 µM) 50 µl
FeCl3 (50 µM) 0.1 ml
ZnCl2 (1 µM) 0.1 ml
thiamine (2 µM) 0.1 ml
0.5% glycerol 0.57 ml
0.5% glutamate 10 ml
tryptophan 1 ml