10 minutes after addition of biotinylated probe and nuclear extract and incubated for an additional 20 minutes at room temperature. Reactions were then loaded onto an 8% TBE gel in 22.25% Tris, pH 8.4, 22.25% boric acid, 0.5 mM EDTA and electrophoresed at 22°C. DNA was transferred to a positively charged nylon membrane, UV cross-linked, probed using Lightshift chemiluminescent EMSA reagents, and detected on a Kodak Image Station 440CF.
