How to Cast Polyacrylamide Gels with the TurboMix™ Bis-Tris Gel Casting Kit

Assemble the gel casting stand and cassette, ensuring glass plates are free of chips and cleaned with alcohol and lint-free wipes. Mark the desired resolving gel height on the plates, typically one centimeter below the comb teeth.

Prepare a fresh 10% APS (ammonium persulfate) solution, which is used in combination with TEMED to initiate and accelerate acrylamide polymerization. APS can be made fresh or aliquoted and stored at -20°C.

Dilute the 20% resolving solution with deionized water to achieve the desired acrylamide percentage (8-15%). For a 10% gel, combine equal volumes of resolving solution and deionized water.

Add TEMED and APS to both the resolving gel (3 µL TEMED and 30 µL APS) and stacking gel (2 µL TEMED and 20 µL APS). Gently invert conicals to mix while avoiding air bubbles, as polymerization will begin immediately.

Pour the resolving gel into the cassette up to the marked height, then immediately add the stacking gel on top without waiting for polymerization. Fill with stacking gel to the top of the cassette to prevent bubble formation around the comb teeth.

Insert the comb at an angle to prevent air from becoming trapped under the teeth. Allow the gel to polymerize in the casting stand for one hour before use or storage.