Team:Goettingen/Project/Materials
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==== 5x M9 salts stock solution ==== | ==== 5x M9 salts stock solution ==== | ||
Stir until dissolved; sterilize by autoclaving at 121°C. <br> | Stir until dissolved; sterilize by autoclaving at 121°C. <br> | ||
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!Amount!Substance | !Amount!Substance | ||
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Revision as of 14:13, 22 September 2012
Deutsch / English |
Contents |
Recipes
Media
Recipes for liquid and solid media for bacterial cultivation. Stir until dissolved; sterilize by autoclaving 121°C. Do not forget a magnetic stirrer.
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
M9 Swimming Agar
- 1.25% (v/v) glycerol
- 20% (v/v) 5x M9 salt stock solution
- 0.1% (v/v) of CaCl2 - 2H2O stock solution (20 mg/mL)
- 0.1% (v/v) MgSO4 stock solution (0.12 g/mL)
- 0.3 % (w/v) of agar
Tryptone swimming agar
- 1% (w/v) bacto tryptone
- 0.5% (w/v) NaCl
- 0.3% (w/v) agar
Antibiotics
Stock solutions and concentrations of antibiotics.
Supplements
- Aminoacid mix (containing all aminoacids except tryptophane): stocksolution 0.2% (w/v) varying amounts used in swimming assays, view methods and notebook of group 1
- Caffeine: stocksolution 10 mM 100 µl applied to whatmanpaper in swimming assays, view methods
- D-Aspartate: stocksolution 10 mM
- 2-Ethyl-1-Hexanole: stocksolution 5 mM 100 µl applied to whatmanpaper in swimming assays, view methods
- Geraniol: stocksolution 5 mM 100 µl applied to whatmanpaper in swimming assays, view methods
- L-Aspartate-4-Benzyl: stocksolution 10 mM 100 µl applied to whatmanpaper in swimming assays, view methods
- L-aspartate: stocksolution 10 mM varying amounts used in swimming assays, view methods and notebook of group 1
- Leucin: stocksolution 30.49 mM add to M9-agar: 4 ml (final concentration 0.305 µM)
- Methionin: stocksolution 26.8 mM add to M9-agar: 4 ml (final concentration 0.268 µM)
- Sodium Cyclamate: stocksolution 10 mM 100 µl applied to whatmanpaper in swimming assays, view methods
- Tryptone: stocksolution 0.5% (w/v) varying amounts used in swimming assays, view methods and notebook of group 1
- Vanillin: stocksolution 10 mM 100 µl applied to whatmanpaper in swimming assays, view methods
Buffer and Solution
Recipes for stocks of buffers and solutions.
5x M9 salts stock solution
Stir until dissolved; sterilize by autoclaving at 121°C.
Amount!Substance |
---|
Na2HPO4 × 7 H2O |
KH2PO4 |
NaCl |
NH4Cl |
ddH2O |
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 |
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
E. coli strain list - List of the working E. coli strains including genotypes, genetic markers, and alleles.
Storage
Storage of E. coli strains - How to keep E. coli strains.
- 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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