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)