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Team:Bielefeld-Germany/Labjournal/Cultivation
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$(".media_1").toggle(); | $(".media_1").toggle(); | ||
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<br><b><u>M9 medium</u></b> | <br><b><u>M9 medium</u></b> | ||
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<p>For 250 mL M9 medium you need 175 mL sterile water (for plates add 4 g Agar-Agar as well). Then add (in the following order:</p> | <p>For 250 mL M9 medium you need 175 mL sterile water (for plates add 4 g Agar-Agar as well). Then add (in the following order:</p> | ||
<p> - 250 µL 100 mM CaCl2 </p> | <p> - 250 µL 100 mM CaCl2 </p> | ||
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<p> - antibiotic stock solution </p> | <p> - antibiotic stock solution </p> | ||
<p> - fill up to 250 mL with sterile water</p> | <p> - fill up to 250 mL with sterile water</p> | ||
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| - | + | <a name="buffer"><span style="color:#ff6600">[Buffers, Solutions]</span></a> | |
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<br><b><u>TAE buffer</u></b> | <br><b><u>TAE buffer</u></b> | ||
<p>For 1 L of 50 x TAE buffer you need:</p> | <p>For 1 L of 50 x TAE buffer you need:</p> | ||
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<p><i>10 mL of the stock is diluted in 1 L dH2O for the gel electrophoresis (0.5 x TAE buffer) </i></p> | <p><i>10 mL of the stock is diluted in 1 L dH2O for the gel electrophoresis (0.5 x TAE buffer) </i></p> | ||
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Revision as of 16:00, 4 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 250 mL M9 medium you need 175 mL sterile water (for plates add 4 g Agar-Agar as well). Then add (in the following order:
- 250 µL 100 mM CaCl2
- 2.5 mL trace salts
- store this stock solution in the dark
- 250 µL MgSO4
- 250 µL 50 mM FeCl3 / 100 mM citrate (one solution, citrate is iron carrier)
- store this stock solution cold and in the dark
- carbon source stock solution (e.g. glucose)
- 50 mL 5x M9 salts stock solution
- 64 g L-1 Na2HPO4 * 7 H2O
- 15 g L-1 KH2PO4
- 2.5 g L-1 NaCl
- 5 g L-1 NH4Cl
- antibiotic stock solution
- fill up to 250 mL with sterile water
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)
