Bio-Rad Mini-PROTEAN® TGX™ Precast Gels for 2-D Electrophoresis - Manual (Page 27)

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Instruction Manual and Application Guide
8.3 Denaturing Nucleic Acid PAGE Buffers
See Appendix B for buffer formulations. Do not adjust pH unless directed to do so.
Running buffer (1x) 89 mM Tris, 89 mM boric acid, 2 mM EDTA
Dilute 100 ml 10x stock (catalog #161-0733) with 900 ml diH
2
O
Sample buffer (5x) 89 mM Tris, 89 mM boric acid, 2 mM EDTA, pH 8.0, 12% Ficoll,
(catalog #161-0768) 0.01% bromophenol blue, 0.02% xylene cyanole FF, 7 M urea
8.4 Sample Preparation
Determine the desired ssDNA or RNA concentration for your sample based on the detection method
used. Dilute 4 parts sample with 1 part sample buffer.
8.5 Running Conditions
Table 8.1. Running conditions for denaturing nucleic acid PAGE with one (1) Mini-PROTEAN TBE-urea gel in the
Mini-PROTEAN Tetra cell. Run conditions and times are approximate and assume a constant voltage of 200 V. When running
more than one gel, current will differ.
10% Gels 15% Gels
Power conditions 200 V constant 200 V constant
Expected current (per gel)
Initial 15 mA 15 mA
Final 10 mA 10 mA
Run time 45–60 min 60–75 min