Bio-Rad Whole Gel Eluter and Mini Whole Gel Eluter User Manual

Page 19

6.3 Separating Gels - Calculating %T

Monomer Concentration

(% T/2.67% C)

%T = x(%)

Acrylamides/Bis - 30% T

c ml

or for 40% T Stock*

or c’ ml

Deionized water

d ml

1.5 M Tris-HCl, pH 8.8

2.5 ml

10% Ammonium Persulfate
(make fresh daily)

50 µl

TEMED

5 µl

Total Monomer

10 ml (volume needed for

two mini gels)

Determine (c ml) and (d ml) for 10 ml of total monomer:

c ml:

Calculate the volume of 30% (or 40%) Acrylamide/Bis stock required for 10 ml
of the desired total monomer concentration (x) with the following formula: For
30% stock:

c ml = (x%)(10 ml)/(30%) = (x)(0.333) ml

ml:

*Similarly, for 40% stock solution:

c' ml = (x%)(10 ml)/(40%) = (x)(0.25) ml

d ml:

Calculate the volume of water required at the desired total monomer concentra-
tion 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 multiplying the
10 ml recipe by the appropriate multiplying factor.

6.4 Stacking Gel - 4% T (0.125 M Tris, pH 6.8)