with the gel 5. when the appropriate amount of separating gel solution has been added (in the case of the Mini-Gel, about 1.5cm from top of front plate or 0.5cm below level where teeth of comb will reach, Fig 1), gently layer about lcm of water on top of the separating gel solution. This keeps the gel surface flat 6. Allow gel to polymerize(30-60 minutes). When the gel has polymerized, a distinct interface will appear between the separating gel and the water, and the gel mold can be tilted to verify polymerization 5cm umpc, menz Fig l Separating gel prior to polymerization Pouring the stacking gel Example of Stacking Gel Preparation Stacking gel (3%): 0.8ml TEMED 0.01ml D(fresh) 004ml TEMED or d was added last 1. Pour off water covering the separating gel. The small droplets remaining will not disturb the stacking gel 2. Combine Solution C and B and water in a small Erlenmeyer flask or a test tube. 3. Add ammonium persulfate and TEMEd and mix by gently swirling or inverting the198 with the gel. 5. When the appropriate amount of separating gel solution has been added (in the case of the Mini-Gel, about 1.5cm from top of front plate or 0.5cm below level where teeth of comb will reach, Fig 1), gently layer about 1cm of water on top of the separating gel solution. This keeps the gel surface flat. 6. Allow gel to polymerize (30-60 minutes). When the gel has polymerized, a distinct interface will appear between the separating gel and the water, and the gel mold can be tilted to verify polymerization. Fig 1. Separating gel prior to polymerization. *Pouring the Stacking Gel Example of Stacking Gel Preparation Stacking gel (3%): B 2.0ml C 0.8ml H2O 5.2ml TEMED 0.01ml D(fresh) 0.04ml Total 8.05ml ⚫ TEMED or D was added last. 1. Pour off water covering the separating gel. The small droplets remaining will not disturb the stacking gel. 2. Combine Solution C and B and water in a small Erlenmeyer flask or a test tube. 3. Add ammonium persulfate and TEMED and mix by gently swirling or inverting the