Team:Goettingen/Project/Materials

Recipes

Media

1. LB-Medium
1% (w/v) bacto tryptone
0.5% (w/v) bacto yeast extract
1% (w/v) NaCl
if required: ad 1% (w/v) agar


2. M9-Medium


3. 0.3% Tryptone swimming agar
1% (w/v) bacto tryptone
0.5% (w/v) NaCl
0.3% (w/v) agar

Antibiotics

Supplements

Buffer and Solution

1. 5x M9 salts stock solution
Stir until dissolved; sterilize by autoclaving at 121°C.


Amount	Substance
64 g	Na2HPO4 × 7 H2O
15 g	KH2PO4
2.5 g	NaCl
5.0 g	NH4Cl
ad 1000 ml	ddH2O




2. CaCl₂ Buffer for competent cells
Stir until dissolved; sterilize by sterile filtration.


Amount	Substance	Notes
7.351 g	CaCl2 × 7 H2O	Alternative 0.1 M CaCl2.
15%	Glycerol
ad 500 ml	ddH2O




3. 50x TAE for agarose gels
Stir until dissolved. Working concentration is 1x (40 ml of 50x stock ad 2 l ddH₂O).


Amount	Substance
242 g	Tris base
57.1 ml	Glacial acetic acid
18.6 g	EDTA
ad 1000 ml	ddH2O

E. coli

Strains

Storage

1. Glycerol Stock
• 900 µl over night culture in LB-liquid medium
• 900 µl 50% sterile glycerol
Glycerol is extremely viscous. Hence, use cut off tips to pipette. Glycerol stocks were vortexed to ensure uniformly mixture and shock frozen in N₂ or dry ice. Then, cultures were stored at -80°C. Do NOT forget to fill in the according GMO S1 construction sheet!

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