小中大
可是我用30mA怎么2.5h时只刚刚进入分离胶呢？
下面是我的配方。麻烦有经验的前辈帮我分析一下。
Separating gel monomer (49.5% T 6% C)
Stacking gel monomer (49.5% T 3% C)
Anode buffer (0.2 M Tris, pH 8.9)
Cathode buffer (0.1 M Tris, 0.1 M tricine, 0.1% SDS, pH 8.25)
It is not necessary to adjust the pH of this buffer, which should be around 8.25. （这个我没调，也没测pH，不知道这样做对不对）
Gel buffer (3.0 M Tris, 0.3% SDS, pH 8.45)
Stacking gel (4.0% T 3.0% C)
In a 50-ml conical tube, mix 0.8 ml of the stacking gel monomer, 2.5 ml of the gel buffer, and 6.7 ml dH2O. Degas under vacuum 10 to 15 minutes. Add 50 µl of 10% ammonium persulfate and 10 µl TEMED. Swirl gently to mix. Use immediately. Produces 10 ml of stacking gel, sufficient for four minigels.
Separating gel (16.5% T 6.0% C)
In a 50-ml conical tube, mix 10.0 ml of the separating gel monomer, 10.0 ml of the gel buffer, 3.1 ml glycerol, and 6.9 ml dH2O. Degas under vacuum 10 to 15 minutes. Add 100 µl of 10% ammonium persulfate and 20 µl TEMED. Swirl gently to mix. Use immediately. Produces 30 ml of separating gel, sufficient for four minigels.
