Bio-Rad DCode™ Universal Mutation Detection System - Manual (Page 43)

40% Acrylamide/Bis Solutions (1.25x TAE, 6M urea)
Reagent
6% Gel 8% Gel 10% Gel 12% Gel
40% Acrylamide/Bis 6.0 ml 8.0 ml 10.0 ml 12.0 ml
50x TAE 1.0 ml** 1.0 ml** 1.0 ml** 1.0 ml**
Urea* 14.4 g 14.4 g 14.4 g 14.4 g
TEMED 40.0 µl 40.0 µl 40.0 µl 40.0 µl
10% Ammonium persulfate 400.0 µl 400.0 µl 400.0 µl 400.0 µl
Total volume 40.0 ml 40.0 ml 40.0 ml 40.0 ml
Add dH
2
O to 40 ml and mix. Cast the gel immediately after adding the TEMED and
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
Amount Final Concentration
Tris base 242.0 g 2 M
Acetic acid, glacial 57.1 ml 1 M
0.5 M EDTA, pH 8.0 100.0 ml 50 mM
dH
2
O to 1,000.0 ml
Mix. Autoclave for 20–30 minutes. Store at room temperature.
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
2% Bromophenol blue 0.25 ml 0.05%
2% Xylene cyanol 0.25 ml 0.05%
100% Glycerol 7.0 ml 70%
dH
2
O 2.5 ml
Total volume 10.0 ml
Store at room temperature.
1.25x TAE Running Buffer
Reagent
Amount
50x TAE buffer 175 ml
dH
2
O 6,825 ml
Total volume 7,000 ml
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