Team:Bielefeld-Germany/Labjournal/Cultivation

From 2013.igem.org

Revision as of 13:50, 5 September 2013 by AnnaK (Talk | contribs)






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 % glucose

20 % Glucose 20 % (w/v)glucose 200 g/L

For 500 mL stock solution add 100 g glucose to 440 mL water. Autoclave for 15 at 121° C

- 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)