1. Assemble vertical slab gel apparatus using 1mm spacer.
Tip:-the plate should be thoroughly cleaned and dried before use.
2. Seal the glass plates on 3 sides with 1% agarose.
3. Pour the separating gel/ resolving gel mixture to a level of approximately 2.5 cm below the glass plates, gently layer 250μl of TDW over the gel surface. Allow to polymerize.
Tips: prepare the solution freshly each time it is required. As soon as ammonium persulphate is added, the gel should be poured quickly before the acrylamide polymerizes.
4. After polymerization remove water from the top.
5. Pour the prepared 5% stacking gel over the resolving gel/ separating gel.
6. Immediately insert a comb and allow polymerizing the gel.