Gel Electrophoresis Protocols

Agarose Gel
10X TBE Running Buffer
Loading Dyes

Agarose Gel

For 1% Agarose, dissolve 1g agarose into 100 ml 0.5X TBE buffer. For 1.5 % Agarose, dissolve 1.5g agarose into 100 ml 0.5X TBE buffer.
Place Erlenmeyer flask containing agarose in solution into microwave for 30 second intervals, swirling between, repeating until agarose is completely dissolved. Allow the solution to cool for one to two minutes (or cool by holding flask under cool, running water while swirling) before loading ethidium bromide (0.5 µg/ml).

10X TBE Running Buffer
Combine the following:
  • 108.0 g    Tris base 
  • 55.0 g     Boric acid 
  • 9.3 g    Na2EDTA

Add ddH2O to 1 liter.
Adjust pH to 8.3 using either NaOH (more basic) or HCl (more acidic).
Dilute to 0.5X for running standard agarose electrophoresis gels. 

Loading Dyes

When making these loading dyes you will notice that the glycerol is difficult to measure because it is like molasses. Use the graduated marks on the side of your tube to measure and cut the bore of the tip to effectively siphon the glycerol. Vortex after mixing and store at 40C.

  1. Bromophenol Blue
    1. EDTA 50 mM
    2. Glycerol 30% (w/v)
    3. Bromophenol Blue 0.5% (w/v)

  2. Xylene Cyanol
    1. EDTA 50 mM
    2. Glycerol 30% (w/v)
    3. Xylene Cyanol 0.5% (w/v)

  3. Bromophenol Blue Xylene Cyanol Mix
    1. EDTA 50 mM
    2. Glycerol 30% (w/v)
    3. Bromophenol Blue 0.25% (w/v)
    4. Xylene Cyanol 0.25% (w/v)