Components of Agarose Gel Electrophoresis

• Sulfonated polysaccharide polymer from the seaweed (genidium spp.)

  Agarose Gel
• Agarose gel is a linear polymer of agarobiose consisting of:

  1,3-linked-B-D-galactopyranose

  1,4-linked-3,6-anhydro-a-L-galactipyranose
• Process of making the gel:

  1. Dried powder is dissolved in hot running buffer.
  2. Cooled between 55-65 degrees celcius then poured to a casting tray.
  3. Comb is inserted to create wells for the sample.
  4. Solidifies at 40 degrees celcius.
  5. Add a little buffer so that the gel doesnt dry out and doesnt get ugly.
• Concentration of agarose gel dictates the size of the spaces in the gel (pore size). Higher concentration, the smaller pore size will be (inversely proportional).
• Separation range (size in bp) if the agarose concentration (%) is 0.3?

  5,000 - 60,000
• Separation range (size in bp) if the agarose concentration (%) is 0.6?

  1,000 - 20,000
• Separation range (size in bp) if the agarose concentration (%) is 0.8?

  800 - 10,000
• Separation range (size in bp) if the agarose concentration (%) is 1.0?

  400 - 8,000
• Separation range (size in bp) if the agarose concentration (%) is 1.2?

  300 - 7,000
• Separation range (size in bp) if the agarose concentration (%) is 1.5?

  200 - 4,000
• Separation range (size in bp) if the agarose concentration (%) is 2.0?

  100 - 3,000