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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)

Terrific Broth (TB)

Terrific broth (TB)

• 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)

His Trap Buffer

His Trap Buffer

• 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

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

TSS buffer

TSS Buffer

• 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

SDS-PAGE running buffer

SDS-PAGE running buffer

• For 1 L:
• 3 g Tris
• 14.4 g Glycine
• 1 g SDS

Colloidal Coomassie Brilliant Blue staining solution

Colloidal Coomassie Brilliant Blue staining solution

• 2.5 g/L Coomassie Brilliant Blue R250
• 10 % (v/v) Acetic Acid
• 25 % (v/v) Isopropyl alcohol

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