Team:Bielefeld-Germany/Labjournal/Cultivation

From 2013.igem.org

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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>
   
   
-
+
 
 +
<br><b><u>M9 medium</u></b>
 +
 
 +
<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> - 2.5 mL trace salts </p>
 +
<p> - store this stock solution in the dark </p>
 +
<p> - 250 µL MgSO4 </p>
 +
<p> - 250 µL 50 mM FeCl3 / 100 mM citrate (one solution, citrate is iron carrier) </p>
 +
<p> - store this stock solution cold and in the dark </p>
 +
<p> - carbon source stock solution (e.g. glucose)</p>
 +
<p> - 50 mL 5x M9 salts stock solution </p>
 +
<p> - 64 g L-1 Na2HPO4 * 7 H2O </p>
 +
<p> - 15 g L-1 KH2PO4</p>
 +
<p> - 2.5 g L-1 NaCl </p>
 +
<p> - 5 g L-1 NH4Cl </p>
 +
<p> - antibiotic stock solution </p>
 +
<p> - fill up to 250 mL with sterile water</p>
 +
 
 +
 
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Revision as of 15:54, 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


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)


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