5% Polyacrylamide gel (30 ml)
22 ml water
3 ml 5X TBE
5 ml 30% Acrylamide
0.210 ml 10% Ammonium persulfate
10-100 ml TEMED
5X TBE (1 L):
54 g Tris base
27.5 g boric acid
20 ml 0.5 M EDTA (pH 8.0)
Reaction:
Combine the components in the following order (in ml):
5X EMSA buffer: 4
Water: to a final volume of 20 ml
PolydI/dC: 1
BSA: 1
32P probe: 1 (10K cpm)
protein: 1-3 ml
let the reaction stand for 10-15 min at room temp., then load 18 ml per lane on a 5% polyacrylamide gel. Run at 150 V for 2h at room temperature, then dry the gel and expose 4-16 h to film at –80 C.
