Bio-Rad Ready Gel Precast Gels for 2-D Electrophoresis - Manual (Page 27)

9.4 TBE-Urea Buffers
Running Buffer Working Concentration 10x Stock
89 mM Tris Tris base 54.0 g
89 mM boric acid Boric acid 27.5
2 mM EDTA 0.5 M EDTA (pH 8.0) 20.0 ml
to 500 ml with deionized water
Sample Buffer Working Concentration
89 mM Tris, 89 mM boric acid, 2 mM EDTA, pH 8.0
10x TBE 1.0 ml
12% Ficoll Ficoll 1.2 g
0.01% Bromophenol Blue Urea 4.2 g
0.02% Xylene Cyanole FF 1% Bromophenol blue 0.1 ml
7 M urea 1% Xylene Cyanole FF 0.2 ml
0.5 M EDTA 0.02 ml
Make up to 10 ml with deionized water
9.5 Sample Preparation
Determine the desired ssDNA or RNA concentration for your sample based on the detection method used.
(See section 10.6 for appropriate stain sensitivities.) Dilute 1 part sample with 1 part TBE-urea sample buffer.
Dry samples can be dissolved directly in sample buffer. Heat to 70–90°C 4 min before loading.
9.6 Running Conditions
Power conditions 200 V constant
Starting current:
15 mA/gel
Final current: 10 mA/gel
Run time 40–70 min
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