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Team:Groningen/protocols/biofilm media
From 2013.igem.org
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<th>compound</th> | <th>compound</th> | ||
Revision as of 08:50, 19 September 2013
The strains were grown overnight at 37°C on LB broth.
Next, 1ml of liquid culture was used to inoculate every 10ml MSgg medium. this in turn was incubated at 27 C for 5 days.
Media for macrocolony formation
2x SG medium (100mL)
| compound | amount | treatment |
|---|---|---|
| Nutrient broth (Difco) | 1.6g | |
| KCl | 0.2g | |
| MgSO47H2O | 0.05g | |
| agar | 1.5g | |
| add al together and autoclave | ||
| Ca(NO3)24H2O 1mM | 0.1mL | |
| MnCl24H2O 0.1mM | 0.1mL | |
| FeSO4 1 uM | 0.1mL | |
| 0.1% glucose | 0.5mL |
LB/Mn/glucose medium (100mL)
| compound | amount | treatment |
|---|---|---|
| LB/ 1.5% agar | 100ml | autoclave |
| MnCl24H2O 0.1mM | 0.1mL | |
| 0.1% glucose | 0.5mL |
MSgg medium (100mL)
| compound | amount | treatment |
|---|---|---|
| KH2PO4 | 0.026g | |
| K2HPO4 | 0.061g | |
| MOPS 100mM | 2.09g | |
| MgCl26H2O 2mM | 0.04g | |
| agar | 1.5g | |
| add al together and autoclave | ||
| CaCl2 700mM | 0.1mL | |
| MnCl2 50uM | 50uL | |
| FeCl3 50 uM | 0.1mL | |
| ZnCl2 1 uM | 0.1mL | |
| thiamine 2 uM | 0.1mL | |
| 0.5% glycerol | 0.57mL | |
| 0.5% glutamate | 10mL | |
| tryptophan | 1mL |
Grow cells overnight in LB / glass tubes at 37C.
Inoculate 2uL spot on plate dry 15 minutes at 30C
Inoculate 30C 2-4 days
iGEM 2013 Groningen
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