Recipes

Lower Gel Buffer (separating)
Tris Base 187 g
SDS 4 g
Bring pH to 8.8 w/HCl
Bring volume up to 1 L with dH2O, filter and store at 4 °C

 

Upper Gel Buffer (stacking)
Tris Base 60.5 g
SDS 4 g
Bring pH to 6.8 w/HCl
Bring volume up to 1 L with dH2O, filter and store at 4 °C

 

Running Buffer
Tris Base 60.6 g
Glycine 288.2 g
SDS 20 g
Bring volume up to 20 L with dH2O

 

5x Sample Buffer
2 M Tris, pH 6.8 1.58 mL
SDS 1 g
2-Mercaptoethanol 2.5 mL
Glycerol 5 mL (1.26 g/mL)
Bromophenol Blue small pinch
Bring volume to 10 mL with dH2O

 

Separating Gel Recipes (for Bio-Rad mini gel)
  7.5% 10% 12.5% 14% 15%
30% Acrylamide/Bis (37.5:1) 2.5 mL 3.3 mL 4.2 mL 4.65 mL 5.0 mL
dH2O 5.0 mL 4.2 mL 3.3 mL 2.85 mL 2.5 mL
Lower Gel Buffer 2.5 mL 2.5 mL 2.5 mL 2.5 mL 2.5 mL
10% APS 75 µL 75 µL 75 µL 75 µL 75 µL
Temed 10 µL 10 µL 10 µL 10 µL 10 µL

 

Stacking Gel
30% Acrylamide/Bis (37.5:1) 0.75 mL
dH2O 3.0 mL
Upper Gel Buffer 1.25 mL
10% APS 50 µL
Temed 10 µL

 

Coomassie Stock
Coomassie R-250 0.15 g
dH2O 100 mL

 

Stain 1 (1 L)
Acetic Acid 100 mL
Isopropanol 100 mL
Coomassie Stock 20 mL
dH2O 780 mL

 

Stain 2 (1 L)
Acetic Acid 100 mL
Coomassie Stock 20 mL
dH2O 880 mL

 

Destain (1 L)
Acetic Acid 100 mL
dH2O 900 mL

 

Gel Soak (1 L)
95% Ethanol (treat as 100%) 300 mL
Glycerol 10 mL
dH2O  

Media

LB-Lennox (1 L)
Tryptone 10 g
Yeast Extract 5 g
dH2O Bring to 1 L

 

Nutrient Broth Soft Agar (200 mL)
Nutrient Broth 1.6 g
NaCl 1.0 g
Agar 1.3 g
dH2O 200 mL

 

SOC (100 mL)
Tryptone 2 g
Yeast Extract 0.5 g
NaCl 50 mg
MgCl2 0.2 g  (or 1 mL 1 M MgCl2)
KCl 18.6 mg  (or 0.25 mL 1 M KCl)
dH2O 100 mL
Glucose (add after autoclaving) 2 mL of 1 M Glucose (sterile filtered)