ReadyPour™ – Protocol
ReadyPour™ Acrylamide Pre-Mix solutions are ready to use. The gel solution contains all components, simply aliquot the desired quantity and add ammonium persulfate and TEMED.
Suggested protocol to cast one gel:
- For the separator gel, aliquot 10 ml of your selected acrylamide percentage and transfer in a clean tube or beaker.
- For the stacking gel, aliquot 2.5 ml of IG001 Stacking solution and transfer in a clean tube.
- Take an empty cassette and place it in an upright position on a support.
- Add 10 μl of TEMED to the Separator solution and mix by swirling a few times.
- Then add 50 μl of 10% APS and mix by swirling a few times.
- Use a pipette to gently deliver the solution to the empty cassette, taking care not to introduce bubbles. The level of solution should just reach 5 to 10 mm from the top of the notched plate.
- Add 2.5 μl of TEMED and 12.5 μl of 10% APS to the Stacking gel solution and swirl to mix.
- Gently add the stacking solution on the top of the gel.
Put the comb in place, and let polymerize at least 10 minutes.
- This protocol has been calibrated for RT defined as 20˚-24˚C, at higher temperatures, the polymerization will be faster. Following this protocol, you have about 2 minutes to pour the gels. This can be difficult if you want to pour several gels simultaneously. However, it is easy to slow the polymerization time: simply reduce the amount of APS 10% to 75 µl, to get about 5 minutes before polymerization. Reduce APS 10% to 50 µl for about 10 minutes of polymerization. Be aware that the stacking gel solution is slower to polymerize than the separator gel solution.