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Casting 12 BioRad Mini SDS PAGE Gels 1. Assemble multicasting chamber. Make sure ALL plates and spacers are flush with bottom. Clean
 

Summary: 120
Casting 12 BioRad Mini SDS PAGE Gels
1. Assemble multicasting chamber. Make sure ALL plates and spacers are flush with bottom. Clean
gasket and coat with a medium-thin layer of Vasoline to avoid leaks.
2. Set up gradient maker and peristaltic pump. Make the 8% and 16% gel solutions in each side of
gradient maker using two stir bars for mixing. No need to degas. Make fresh ammonium persulfate
(25% APS = 0.25 g APS + 0.85 ml ddH2O). Make sure valve and tube to chamber are not clogged.
8% 16%
5X 8.8 Buffer 5.5 ml 5.5 ml
30% Acrylamide 7.5 ml 14.5 ml
ddH2O 14.5 ml 2 ml
50% Glycerol -- 5.5 ml
TEMED 28 Ál 28 Ál
25% APS 35 Ál 35 Ál
3. After adding APS with stirring, open end and middle valves. Pump at speed "8" to pour gels (~10
min). Stop pumping when resolving gel is 1 cm below where comb teeth will be. Do not pump air
past valve at bottom of multicasting chamber. Overlay each gel with 0.15 ml butanol (ddH2O sat).
4. After polymerization (~30 min), wash with ddH2O 3-5 times. Dry with triangles of filter paper.
5. Prepare stacking gel. Degas with vacuum aspirator. Use APS made above.
4.5%

  

Source: Aris, John P. - Department of Anatomy and Cell Biology, University of Florida

 

Collections: Biology and Medicine