BIO RAD DCODE Manual page 41

40% Acrylamide/Bis Solutions (1.25x TAE, 6M urea)
Reagent
40% Acrylamide/Bis
50x TAE
Urea*
TEMED
10% Ammonium persulfate
Total volume
Add dH O to 40 ml and mix. Cast the gel immediately after adding the TEMED and
2
ammonium persulfate.
* For 7 M urea gels, use 16.8 g per 40 ml, for 8 M urea gels, use 19.2 g per 40 ml.
** Adjust this amount for other concentrations of running buffer.
50x TAE Buffer
Reagent
Tris base
Acetic acid, glacial
0.5 M EDTA, pH 8.0
dH O
2
Mix. Autoclave for 20–30 minutes. Store at room temperature.
10% Ammonium Persulfate
Reagent
Ammonium persulfate
dH O
2
Store at -20 °C for about a week.
2x Gel Loading Dye
Reagent
2% Bromophenol blue
2% Xylene cyanol
100% Glycerol
dH O
2
Total volume
Store at room temperature.
1.25x TAE Running Buffer
Reagent
50x TAE buffer
dH O
2
Total volume
6% Gel 8% Gel
6.0 ml 8.0 ml
1.0 ml** 1.0 ml**
14.4 g 14.4 g
40.0 µl 40.0 µl
400.0 µl 400.0 µl
40.0 ml 40.0 ml
Amount
242.0 g
57.1 ml
100.0 ml
to 1,000.0 ml
Amount
0.1 g
1.0 ml
Amount
0.25 ml
0.25 ml
7.0 ml
2.5 ml
10.0 ml
Amount
175 ml
6,825 ml
7,000 ml
37
10% Gel 12% Gel
10.0 ml 12.0 ml
1.0 ml** 1.0 ml**
14.4 g 14.4 g
40.0 µl 40.0 µl
400.0 µl 400.0 µl
40.0 ml 40.0 ml
Final Concentration
2 M
1 M
50 mM
Final Concentration
0.05%
0.05%
70%