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Team:Bielefeld-CeBiTec/Notebook/Media
From 2014.igem.org
Media
- 12 g Tryptone
- 24 g yeast extract
- 4 ml Glycerol
- dissolve the solutes in 900 ml of deionized H2O
- Salt Solution:
- KH2PO4 (0,17M) equals 2.31 g per 100 milliliters
- K2HPO4 (0,72 M) equals 12.54 g per 100 millilters
- dissolve the solutes in 100 ml of deionized H2O and mix them with the other media components to get a total volume of 1 litre
- 15 g Agar-Agar per Liter (for plates)
- All buffers with pH 7.4 - 7.6
| Name | Sodium phosphate | NaCl | Imidazol | EDTA |
|---|---|---|---|---|
| Binding Buffer | 20 mM | 500 mM | 5 mM | 0 mM |
| Elution Buffer 1 | 20 mM | 500 mM | 40 mM | 0 mM |
| Elution Buffer 2 | 20 mM | 500 mM | 60 mM | 0 mM |
| Elution Buffer 3 | 20 mM | 500 mM | 100 mM | 0 mM |
| Elution Buffer 4 | 20 mM | 500 mM | 300 mM | 0 mM |
| Elution Buffer 5 | 20 mM | 500 mM | 500 mM | 0 mM |
| Stripping Buffer | 20 mM | 500 mM | 0 mM | 50 mM |
- To prepare 1 liter of M9 minimal medium add the following components to 836.7 mL sterile distilled water. To avoid precipitation, start with CaCl2 and mix the solution thoroughly after addition of a component.
- 100 mL 10 X M9 salt solution
- 50 mL 20 X carbon source
- 1 mL 1000 X trace element solution
- 1 mL autoclaved 1 M MgSO4
- 0.3 mL autoclaved 1 M CaCl2
- 1 mL filter sterilized 1 g/L biotin
- 1 mL filter sterilized 1 g/L thiamin
- 75.2 g Na2HPO4 x 2H2O
- 30 g KH2PO4
- 5 g NaCl
- 5 g NH4Cl
- Dissolve the salts in 800 mL water and adjust the pH to 7.2 with NaOH. Add water to a final volume of 1 L and autoclave for 15 minutes at 121 °C.
- Add the following components for 900 ml of distilled H2O:
- 20 g Trypton
- 5 g Bacto Yeast Extract
- 2 mL of 5 M NaCl
- 2.5 ml of 1 M KCl
- 10 ml of 1 M MgCl2
- 10 ml of 1 M MgSO4
- 20 ml of 1 M glucose
- For each litre of solution:
- 242 g Tris Base (MW=121.1)
- 57.1 mL Glacial Acetic Acid
- 100 mL 0.5 M EDTA
- mix Tris with stir bar to dissolve in about 600 mL of ddH2O.
- add the EDTA and Acetic Acid.
- bring final volume to 1 L with ddH20.
- store at room temperature.
- Note: Final (1x) working concentration:
- 0.04 M Tris - Acetate
- 0.001 M EDTA
- For each litre of solution:
- 242 g Tris Base (MW=121.1)
- 57.1 mL Glacial Acetic Acid
- 10 mL 0.5 M EDTA
- mix Tris with stir bar to dissolve in about 600 mL of ddH2O.
- add the EDTA and Acetic Acid, pH to 8.0.
- bring final volume to 1 L with ddH2O.
- store at room temperature.
- Note: Final (1x) working concentration:
- 0.04 M Tris - Acetate
- 0.0001 M EDTA
- 1 L of 50x TAE buffer
- 242.48 g Tris
- 41.02 g sodium acetate
- 18.612 g EDTA
- Adjust pH to 7.8 with acetic acid
- Solve in dH2O
- Dilute 20 mL 50x stock in 1L dH2O for 1x Buffer for PAGE
- 292.243g/mol 1mM EDTA
- 0.025g 0.05% (w/v) BPB
- 0.025g 0.05% (w/v) Xylene Cyanol
- Solve in H2O
- Adjust color to green with HCl
- Dilute with glycerol to 50:50
- For 50 mL:
- 5g PEG 8000
- 1.5 mL 1M MgCl2 (or 0.30g MgCl2*6H2O)
- 2.5 mL DMSO
- Add LB to 50 mL
- Store at 4°C or -20°C
- 2.5 g/L Coomassie Brilliant Blue R250
- 10 % (v/v) Acetic Acid
- 25 % (v/v) Isopropyl alcohol
- 3mL of 1M Tris-HCl (pH 7.5)
- 150 µL of 2 M MgCl2
- 60 µL of 100 mM dGTP
- 60 µL of 100 mM dATP
- 60 µL of 100 mM dTTP
- 60 µL of 100 mM dCTP
- 300 µL of 1 M DTT
- 1.5 g PEG-8000
- 300 µL of 100 mM NAD
