Team:Bielefeld-CeBiTec/Notebook/Media

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Media

LB medium

LB medium

  • 10 g Trypton
  • 5 g yeast extract
  • 10 g NaCl
  • 12 g Agar-Agar (for plates)
M9 medium

M9 medium

  • 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
M9 salt stock solution

M9 salt stock solution

  • 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.
SOC-medium

SOC-medium

  • 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
50X TAE Stock Solution

50X TAE Stock Solution

  • 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
50X Modified TAE Stock Solution

50X Modified TAE Stock Solution

  • 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
TAE buffer

TAE buffer

  • 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
Loading buffer

Loading Buffer

  • 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
5x isothermal reaction buffer for Gibson Assembly

5x isothermal reaction buffer for Gibson Assembly

  • 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

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