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Using Acrylamide and Bis-acrylamide Stock Solutions
Use these calculations to determine the necessary volumes of stock acrylamide and
bis-acrylamide solutions to produce gels of any percent and volume.
To determine the monomer and crosslinker ratios:
1.
(A) is the part of total monomer that is acrylamide
A =
gm acrylamide
gm acry gm bis-acrylamide
2.
(B) is the part of total monomer that is bis-acrylamide
B =
gm bis-acrylamide
gm acry gm bis-acrylamide
To determine the volume of 40% acrylamide stock solution to use:
3.
Use the calculation for (A) determined above.
(A) (gel %) (final volume)
= ml of 40% acrylamide (C)
(40%) acrylamide solution
4.
Use the calculation for (B) determined above.
(B) (gel %)(final volume)
= ml of 2% bis-acrylamide (D)
(2%) bis-acrylamide solution
Acrylamide/Bis Solutions (1x TBE)
Reagent
8% Gel (37.5:1)
X% Gel
40% Acrylamide
7.78 ml
(C) from above
2% Bis
4.27 ml
(D) from above
10x TBE (see note)
4 ml
4 ml
100% Glycerol (optional)
see note
see note
TEMED
40 µl
40 µl
10% Ammonium persulfate
400 µl
400 µl
dH
2
O
to 40 ml
to 40 ml
Cast the gel immediately after adding the TEMED and ammonium persulfate.
Optional: 0.5x TBE, add 2 ml
5% glycerol, add 2 ml
10% glycerol, add 4 ml
10% Ammonium Persulfate
Reagent
Amount
Ammonium persulfate
0.1 g
dH
2
O
1.0 ml
Store at -20 °C for about a week.
2x Gel Loading Dye
Reagent
Amount
Final Concentration
Bromophenol blue
0.05 g
0.05%
Xylene cyanol
0.05 g
0.05%
Formamide
9.5 ml
95%
0.5 M EDTA, pH 8
0.4 ml
20 mM
Total volume
10.0 ml
Store at room temperature.
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Summary of Contents for DCODE
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