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SDS - Polyacrylamide Gels


 Separation gel:

for 10 ml

for 20 ml
Gel % 6.0 7.5 8.0 10 12 6.0 7.5 8.0 10 12
4 x lower buffer (in ml) 2.5 2.5 2.5 2.5 2.5 5.0 5.0 5.0 5.0 5.0
40 % (30 : 0.8) acrylamide/
bisacrylamide (in ml)		 1.5 1.9 2.0 2.5 3.0 3.0 3.8 4.0 5.0 6.0
H2O (in ml) 6.0 5.6 5.5 5.0 4.5 12.0 11.2 11.0 10.0 9.0
APS (25 %) in µl 30 30 30 30 30 60 60 60 60 60
TEMED in µl 15 15 15 15 15 30 30 30 30 30


 5 % stacking gel:

for 5 ml for 10 ml
4 x upper buffer (in ml) 1.25 2.5
40 % (30 : 0.8) acrylamide/
bisacrylamide (in µl)		 625 1250
H2O (in ml) 3.1 6.25
APS (25 %) in µl 30 60
TEMED in µl 15 30



Buffers:

 4 x lower buffer: 1.5 M Tris/HCl pH 8.8; 0.4% SDS.
[for 500ml: 91 g Tris/HCl; 2 g SDS]

 4 x upper buffer: 0.5 M Tris/HCl pH 6.8; 0.4% SDS.
[for 500ml: 30.4 g Tris/HCl; 2 g SDS]

 10 x running buffer: 0.25 M Tris; 2 M Glycin; 1 % SDS.
[for 1 l: 30.4 g Tris; ; 150 g Glycin; 10 g SDS]

 2 x loading buffer (reducing): 80 mM Tris/HCl pH 6.8; 10 % Mercaptoethanol;
2 % SDS; 10 % Glycerin; Bromphenol Blue.
[for 10 ml: 800 µl 1 M Tris/HCl pH 6.8; 1 ml Mercaptoethanol; 2 ml 10 % SDS; 2 ml 50 % Glycerin, Bromphenol Blue]
 
Last Modified: Aug 24, 2000


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