Bio-Rad Mini-PROTEAN 3 Multi-Casting Chamber - Manual (Page 8)

Premixed Gel Casting Buffers
161-0798 Resolving Gel Buffer 1.5 M Tris-HCl , pH 8.8, 1L
161-0799 Stacking Gel Buffer 0.5 M Tris-HCl , pH 6.8, 1 L
Appendix A
Estimated Volume of Acrylamide for
12 Mini-PROTEAN 3 Gels
The first time gels of a certain thickness are cast, it is necessary to empirically determine
the required volume of acrylamide. Assemble the stack, and inject a measured volume of
water through the stopcock. Prepare this volume (+5 ml) of acrylamide.
As a guideline, this is the estimated volume of acrylamide for 12 Mini-PROTEAN 3 gels:
Spacer plates Volume for 12 gels Volume to prepare
0.5 mm 70 ml 75–80 ml
0.75 mm 80 ml 85–90 ml
1.0 mm 100 ml 105–110 ml
1.5 mm 140 ml 145–150 ml
Sample calculation: preparing twelve 1.0 mm 4–20% gradient gels:
Casting twelve 1.0 mm gels requires 100 ml; prepare 110 ml.
Divide the total volume by 2 to get the volume required for each chamber. (For this example,
make 55 ml for the light chamber and 55 ml for the heavy chamber.)
Solution Volume Calculations:
Light Solution (4%)
Acrylamide
30% stock solution
(30%) (X ml) = (4%) (55 ml)
X = 7.3 ml
Tris-Cl Buffer
1.5M Tris-Cl stock buffer pH 8.8
(1.5M) (X ml) = (.375M) (55 ml)
X = 13.8 ml
Water
(55 ml)–(7.3 ml + 13.8 ml) = X
X = 34 ml
APS
10% Stock solution
(500 µl) / (100 ml) = (X µl) / (55 ml)
X = 275 µl
TEMED
10% of the APS volume; (275 µl) / 10 = X
X = 27.5 µl
Heavy Solution (20%)
Acrylamide
30% Stock solution
(30%) (X ml) = (20%) (55 ml)
X = 36.7 ml
Tris-Cl Buffer
1.5M Tris-Cl Stock Buffer pH 8.8
(1.5M) (X ml) = (.375M) (55 ml)
X = 13.8 ml
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