Bio-Rad Mini-PROTEAN® TGX™ Precast Gels for 2-D Electrophoresis - Manual (Page 41)

Technical Support: 1-800-4BIORAD • 1-800-424-6723 • www.bio-rad.com 35
Instruction Manual and Application Guide
Running buffer (1x) Dilute 100 ml 10x stock (catalog #161-0733) with 900 ml diH
2
O.
Sample buffer (5x) 50 mM Tris-HCl, pH 8.0, 5mM EDTA, 25% glycerol, 0.2% bromophenol
(catalog #161-0767) blue, 0.2% xylene cyanole FF
Prepare Buffers
Prepare and Load Samples
Prepare Gels and
Assemble Electrophoresis Cell
Perform Electrophoresis
Remove the comb and tape from the gels and assemble the electrophoresis cell.
Fill the inner and outer buffer chambers with running buffer.
Nondenaturing Nucleic Acid PAGE (Mini-PROTEAN TBE Gels)
Component Amount
Sample 8 μl
Sample buffer
(catalog #161-0767) 2 μl
Total volume 10 μl
Load the appropriate amount of sample on the gel.
Connect the electrophoresis cell to the power supply and perform electrophoresis according to
the conditions in the table.
Table A.4. 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