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Team:Bielefeld-Germany/Labjournal/Cultivation
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<br><b><u>M9 medium</u></b> | <br><b><u>M9 medium</u></b> | ||
<p>For 1 L <b>M9 mineral medium</b> you need 867 mL sterile water <i>(for plates add 16 g Agar-Agar as well)</i>. Then add (in the following order:</p> | <p>For 1 L <b>M9 mineral medium</b> you need 867 mL sterile water <i>(for plates add 16 g Agar-Agar as well)</i>. Then add (in the following order:</p> | ||
| - | <p> - 300 µL 1 M CaCl2 </p> | + | <p> - 300 µL 1 M CaCl2 |
| + | |||
| + | </p> | ||
| + | |||
| + | |||
<p> - 100 mL 5x M9 salt solution</p> | <p> - 100 mL 5x M9 salt solution</p> | ||
| + | <table border=1 style="float:left; padding-left:10px"> | ||
| + | <col width="150"> | ||
| + | <col width="50"> | ||
| + | <tr><th><b> 10x M9 salt solution</b></th><th></th><tr> | ||
| + | <tr> <td align="right">Na2HPO4-2H2O</td> <td align="center">75.2 g/L</td> </tr> | ||
| + | <tr> <td align="right">KH2PO4</td> <td align="center">30 g/L</td> </tr> | ||
| + | <tr> <td align="right">NaCl</td> <td align="center">5 g/L</td> </tr> | ||
| + | <tr> <td align="right">NH4Cl</td> <td align="center">5 g/L</td> </tr> | ||
| + | |||
| + | </table> | ||
| + | <br><br><br><br><br> | ||
<p> - 20 mL 20 % glucose</p> | <p> - 20 mL 20 % glucose</p> | ||
<p> - 1 M MgSO4 </p> | <p> - 1 M MgSO4 </p> | ||
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Revision as of 11:33, 5 September 2013
Cultivation
Chloramphenicol stock solution
-Solubilize 20 mg mL-1 Chloramphenicol in 100 % Ethanol
-Store at -20 °C
DNA loading buffer
- 50 % (v/v) glycerol
- 1 mM EDTA
- 0.1 % (w/v) bromphenol blue
- Solve in ddH2O
LB medium
- 10 g Trypton
- 5 g yeast extract
- 10 g NaCl
- 12 g Agar-Agar (for plates)
- Adjust pH to 7.4
M9 medium
For 1 L M9 mineral medium you need 867 mL sterile water (for plates add 16 g Agar-Agar as well). Then add (in the following order:
- 300 µL 1 M CaCl2
- 100 mL 5x M9 salt solution
| 10x M9 salt solution | |
|---|---|
| Na2HPO4-2H2O | 75.2 g/L |
| KH2PO4 | 30 g/L |
| NaCl | 5 g/L |
| NH4Cl | 5 g/L |
- 20 mL 20 % glucose
- 1 M MgSO4
- 1 mL biotin (1 mg/mL)
- 1 mL thiamin (1 mg/mL)
- 10 mL 100x trace elements solution
TAE buffer
For 1 L of 50 x TAE buffer you need:
- 242.48 g Tris
- 41.02 g Sodiumacetate
- 18.612 g EDTA
- Adjust pH to 7.8 with acetic acid
- Solve in dH2O
10 mL of the stock is diluted in 1 L dH2O for the gel electrophoresis (0.5 x TAE buffer)
