AccuGel 29:1 (40%)

Protocol

Procedure

Mix Gel Solution

Calculate how much AccuGel you need to make your gels by using the formulas below. Bring up to the desired final volume with your usual buffers and distilled water. Pour the solution into an Erlenmeyer flask with a side-arm. In most cases, AccuGel will gel without degassing. However, for optimum reproducibility, add a stirring bar to the solution and stopper the flask. Degas the solution under a vacuum for 5 minutes while stirring on a magnetic stirrer.

where:
V_t = Total volume of gel casting (solution desired (ml))
V_A30 = Volume of 30% AccuGel to be used (ml)
OR
V_A40 = Volume of 40% AccuGel to be used (ml)

Initiate and Cast Gel:

Add 1.0ml of 10% (w/v) FRESHLY PREPARED ammonium persulfate for every 100ml of gel casting solution. Swirl gently to mix. Add 100 microliters of TEMED for every 100ml of gel casting solution. Swirl gently to mix. Pour the solution into the gel casting cassette. The gel should begin to set in 10-20 minutes. Polymerization should be permitted to continue for a minimum of 1.5-2 hours before gel is run.

NOTE: After two hours of polymerization wrap each end of the gel cassette with clear plastic wrap. This is important to keep the ends of the gel from drying and to maintain sample well integrity. Appropriately wrapped gels may be stored for up to 48 hours.

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