Bis-AcrylaGel

Protocol

Procedure

Mix Gel Solution

Calculate how much AcrylaGel and Bis-AcrylaGel 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.

V_a = (A)(V_t)/30
V_b = (A)(C)(V_t)/200

where,

V_a = Volume of AcrylaGel to be used (ml),
V_b = Volume of Bis-AcrylaGel to be used (ml),
V_t = Total volume of gel casting solution desired (ml),
A = % acrylamide desired in gel,
C = % crosslinker desired

EXAMPLE: To make 100ml of a 10% acrylamide gel 2.7% crosslinked with bis, calculate the volume to be added as follows:
V_a = (10) (100)/30 = 33.3ml AcrylaGel
V_b = (10) (2.7) (100)/200 = 13.5ml Bis-AcrylaGel

Add APS 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 0.1ml 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.

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