Bio-Rad Mini-PROTEAN® TGX™ Precast Gels for 2-D Electrophoresis - Manual (Page 25)

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7.3 Nondenaturing 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) 50 mM Tris-HCl, pH 8.0, 5 mM EDTA, 25% (w/v) glycerol, 0.2% bromophenol
(catalog #161-0767) blue, 0.2% xylene cyanole FF
7.4 Sample Preparation
Determine the DNA concentration of your sample based on the detection method used. (See Chapter 10
for approximate stain sensitivities.) Dilute 4 parts sample with 1 part sample buffer.
7.5 Running Conditions
Table 7.1. Running conditions for nondenaturing nucleic acid PAGE with one (1) Mini-PROTEAN TBE gel in the
Mini-PROTEAN Tetra cell. Run conditions and times are approximate and assume a constant voltage of 100 V. When running
more than one gel, current will differ.
5% Gels 10% Gels 15% Gels 4–20% Gels
Power conditions 100 V constant 100 V constant 100 V constant 100 V constant
Expected current (per gel)
Initial 15 mA 15 mA 15 mA 15 mA
Final 10 mA 10 mA 10 mA 10 mA
Run time 45–60 min 60–75 min 75–90 min 90–105 min
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