ACRYLAMIDE GEL PROCEDURE

Gel Cassette Assembly (Bio-Rad Mini Protean 3)
 



Table 1.  Denaturing acrylamide gel conditions

Gel Composition 
6%
8%
10%
12%
Water (mL)
6.4
5.9
5.4
4.9
5X TBE (mL)
2.0
2.0
2.0
2.0
Ureaa (g)
4.8
4.8
4.8
4.8
40% acrylamide (mL)
1.5
2.0
2.5
3.0
10% APS (mcL)
100
100
100
100
TEMED (mcL)
10
10
10
10
Total Volume (mL)
10
10
10
10
                                                                              a the best practice would be to dissolve the urea in a
                                                                                small volume of water, add the buffer and acrylamide
                                                                                then adjust the volume to 10 mL

Table 2.  Native acrylamide gel conditions

Gel Composition 
6%
8%
10%
12%
Water (mL)
6.4
5.9
5.4
4.9
5X TBE (mL)
2.0
2.0
2.0
2.0
40% acrylamide (mL)
1.5
2.0
2.5
3.0
10% APS (mcL)
100
100
100
100
TEMED (mcL)
10
10
10
10
Total Volume (mL)
10
10
10
10

Table 3.  SDS-PAGE gel conditions

Gel Composition 
6%
8%
10%
12%
Water (mL)
6.4
5.9
5.4
4.9
1.5M Tris (mL)
2.6
2.6
2.6
2.6
10% SDS (mcL)
100
100
100
100
40% acrylamide (mL)
1.5
2.0
2.5
3.0
10% APS (mcL)
100
100
100
100
TEMED (mcL)
10
10
10
10
Total Volume (mL)
10
10
10
10


Electrophoresis



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