Owner's manual

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8.2 Analytical Separating Gels
Table 10. Calculating %T (0.375 M Tris, pH 8.8).
Separating Monomer Concentration = %T = %x
Acrylamide/bis (30% T/2.67%C Stock) c ml
Deionized water d ml
1.5 M Tris HCI, pH 8.8 2.5 ml
10% ammonium persulfate (fresh daily) 50 µl
TEMED 5 µl
Total monomer 10 ml (volume needed for 2 mini gels
Determine c ml and d ml for 10 ml of total monomer:
c ml: Calculate the volume of 30% acrylamide/bis stock
required for 10 ml of the desired total monomer concentration
with the following formula:
c ml = (%x)(10 ml)/(30%) = (x)(0.333) ml
d ml: Calculate the volume of water required at the
desired total monomer concentration with the
following formula:
d ml = 10 ml – 2.5 ml – 50 µl – 5 µl – c ml = 7.445 ml – c ml
Important: One can prepare any desired volume of monomer solution by mul tiplying the 10 ml recipe by the
appropriate multiplying factor.
8.3 Analytical Stacking Gel
Table 11. 4%T/2.67%C (0.125 M Tris, pH 6.8).
Stacking Monomer Concentration = 40% T/2.67% C
Acrylamide/bis (30% T/2.67%C stock) 1.33 ml
0.5 M Tris HCI, pH 6.8 6.10 ml
Deionized water 2.5 ml
10% ammonium persulfate (fresh daily) 50 µl
TEMED 10 µl
Total monomer 10 ml (volume needed for 2 stacking gels
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