SDS-PAGE on vertical mini gels:

• In an Erlenmeyer flask, add 1 g of Agarose to 50 ml of water
• Solubilize the 2% agarose solution in a microwave oven for two minutes
• pour the hot solution on a clean bench surface
• assemble the gel box by separating the two glass layers by space fillers and immobilizing them with the provided plastic grips
• lay down the assembled gel box and let the hot solution seal the bottom side
• after a few minutes, fill the inner cavity of the box sparing the top 2-2.5 cm, with ca. 10 ml of the following 10% gel:

  2.5 ml Acrilamide/bisacrilamide 40%
  2.5 ml water
  5.0 ml Tris 0.5M (pH= 8.8, adjusted with HCl)
  80 µl solution SDS 10%
  immediately before the introduction of the gel in the assembled gel box, add the following reagents in this order:
  33 µl solution 25% ammonium persulfate (0.57 g in 10 ml water)
  9 µl TEMED
  
  

• add isobutanol to cover the gel, let it solidify and flush then with deionized water
• to the empty upper part of the solidified gel, add a 5% gel solution prepared as follows:

  1.25 ml Acrilamide/bisacrilamide 40%
  6.25 ml water
  2.5 ml Tris 0.5M (pH= 6.8, adjusted with HCl)
  100 µl solution SDS 10%
  immediately before the introduction of the gel in the assembled gel box, add the following reagents in this order:
  40 µl solution 25% ammonium persulfate (0.57 g in 10 ml water)
  15 µl TEMED
  
  

• introduce the comb and wait for the solidification of the gel
• place the vertical gel in the machine (?) and remove the comb
• fill the upper basket with 250 ml of SDS-PAGE solution
• introduce 15 µl of the sample (with SDS) and start the electrophoresis (140V; 60A)