Team:Groningen/protocols/biofilm media
From 2013.igem.org
(Difference between revisions)
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+ | 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°C for 5 days. | ||
- | < | + | <h5>Media for macrocolony formation</h5> |
- | + | <H5 id="special">2x SG Medium (100 ml)</H5> | |
- | + | <table id="normal" width="70%"> | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | <table id="normal"> | + | |
<tr> | <tr> | ||
<th>compound</th> | <th>compound</th> | ||
Line 42: | Line 37: | ||
<tr> | <tr> | ||
<td>Nutrient broth (Difco)</td> | <td>Nutrient broth (Difco)</td> | ||
- | <td | + | <td>1.6 g</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 48: | Line 43: | ||
<tr> | <tr> | ||
<td>KCl</td> | <td>KCl</td> | ||
- | <td | + | <td>0.2 g</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 54: | Line 49: | ||
<tr> | <tr> | ||
<td>MgSO<sub>4</sub>.7H<sub>2</sub>O </td> | <td>MgSO<sub>4</sub>.7H<sub>2</sub>O </td> | ||
- | <td | + | <td>0.05 g</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 60: | Line 55: | ||
<tr> | <tr> | ||
<td>agar</td> | <td>agar</td> | ||
- | <td | + | <td>1.5 g</td> |
- | <td> </td> | + | <td></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td></td> | <td></td> | ||
- | <td | + | <td></td> |
<td> add everything 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 1 mM</td> | + | <td>Ca(NO<sub>3</sub>)<sub>2</sub>.4H<sub>2</sub>O (1 mM)</td> |
- | <td | + | <td>0.1 ml</td> |
- | <td> </td> | + | <td></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>MnCl<sub>2</sub>.4H<sub>2</sub>O 0.1 mM</td> | + | <td>MnCl<sub>2</sub>.4H<sub>2</sub>O (0.1 mM)</td> |
- | <td | + | <td>0.1 ml</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>FeSO<sub>4</sub> 1 µM</td> | + | <td>FeSO<sub>4</sub> (1 µM)</td> |
- | <td | + | <td>0.1 ml</td> |
- | <td> </td> | + | <td></td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>0.1% glucose </td> | <td>0.1% glucose </td> | ||
- | <td | + | <td>0.5 ml</td> |
- | <td> </td> | + | <td></td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 97: | Line 92: | ||
- | <br><H5>LB/Mn/ | + | <br><H5 id="special">LB/Mn/Glucose Medium (100 ml)</H5> |
- | <table id="normal"> | + | <table id="normal" width="70%"> |
<tr> | <tr> | ||
<th>compound</th> | <th>compound</th> | ||
Line 106: | Line 101: | ||
<tr> | <tr> | ||
- | <td>LB | + | <td>LB 1.5% agar</td> |
- | <td | + | <td>100 ml</td> |
<td>autoclave</td> | <td>autoclave</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>MnCl<sub>2</sub>.4H<sub>2</sub>O 0.1 mM</td> | + | <td>MnCl<sub>2</sub>.4H<sub>2</sub>O (0.1 mM)</td> |
- | <td | + | <td>0.1 ml</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> | ||
Line 119: | Line 114: | ||
<tr> | <tr> | ||
<td>0.1% glucose </td> | <td>0.1% glucose </td> | ||
- | <td | + | <td>0.5 ml</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> | ||
Line 127: | Line 122: | ||
- | <br><H5> MSgg | + | <br><H5 id="special"> MSgg Medium (100 ml)</H5> |
- | <table id="normal"> | + | <table id="normal" width="70%"> |
<tr> | <tr> | ||
<th>compound</th> | <th>compound</th> | ||
Line 137: | Line 132: | ||
<tr> | <tr> | ||
<td>KH<sub>2</sub>PO<sub>4</sub></td> | <td>KH<sub>2</sub>PO<sub>4</sub></td> | ||
- | <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> | ||
- | <td | + | <td>0.061 g</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>MOPS 100 mM</td> | + | <td>MOPS (100 mM)</td> |
- | <td | + | <td>2.09 g</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>MgCl<sub>2</sub>.6H<sub>2</sub>O 2 mM</td> | + | <td>MgCl<sub>2</sub>.6H<sub>2</sub>O (2 mM)</td> |
- | <td | + | <td>0.04 g</td> |
<td></td> | <td></td> | ||
</tr> | </tr> | ||
Line 160: | Line 155: | ||
<tr> | <tr> | ||
<td>agar</td> | <td>agar</td> | ||
- | <td | + | <td>1.5 g</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> | ||
Line 166: | Line 161: | ||
<tr> | <tr> | ||
<td></td> | <td></td> | ||
- | <td | + | <td></td> |
<td>add everything together and autoclave</td> | <td>add everything together and autoclave</td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>CaCl<sub>2</sub> 700 mM</td> | + | <td>CaCl<sub>2</sub> (700 mM)</td> |
- | <td | + | <td>0.1 ml</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>MnCl<sub>2</sub> 50 µM</td> | + | <td>MnCl<sub>2</sub> (50 µM)</td> |
- | <td | + | <td>50 µl</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>FeCl<sub>3</sub> 50 µM</td> | + | <td>FeCl<sub>3</sub> (50 µM)</td> |
- | <td | + | <td>0.1 ml</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>ZnCl<sub>2</sub> 1 µM</td> | + | <td>ZnCl<sub>2</sub> (1 µM)</td> |
- | <td | + | <td>0.1 ml</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td>thiamine 2 µM</td> | + | <td>thiamine (2 µM)</td> |
- | <td | + | <td>0.1 ml</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> | ||
Line 202: | Line 197: | ||
<tr> | <tr> | ||
<td>0.5% glycerol </td> | <td>0.5% glycerol </td> | ||
- | <td | + | <td>0.57 ml</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> | ||
Line 208: | Line 203: | ||
<tr> | <tr> | ||
<td>0.5% glutamate </td> | <td>0.5% glutamate </td> | ||
- | <td | + | <td>10 ml</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> | ||
Line 214: | Line 209: | ||
<tr> | <tr> | ||
<td>tryptophan </td> | <td>tryptophan </td> | ||
- | <td | + | <td>1 ml</td> |
<td> </td> | <td> </td> | ||
</tr> | </tr> |
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.
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 |