Optimized Gel Electrophoresis Visualization

For gel electrophoresis we used 1%-broad-range-agarose (Carl Roth) gels and TAE buffer. We used the 1 kb gene ruler (Thermo Scientific; 5 μl) as a size standard. The gels were run for 120 min at 75 V at 4°C. Subsequently, the gels were removed from the gel box and placed for 20 minutes in 100 ml of 0.5 μM (1:100 000 dilution of the stock; 2 independent gels), 5 μM (1:10 000 dilution of the stock; 7 independent gels), or 50 μM (1:1000 dilution of the stock; 2 independent gels) EtBr in TAE buffer, respectively, for staining. Subsequently, the gel was de-stained in TAE buffer for 15 min. The gels were visualized using a Gel Doc XR+ system (Biorad). The same procedure was employed for SYBR Gold staining, except that the gels were stained for 20 minutes in 100 ml of 3 (1:4000 dilution of the stock; 2 gels in total) or 6 μM (1:2000 dilution of the stock; 2 gels in total). Since the dye concentration used in staining is reduced by the agarose gel matrix and the de-staining step, we used a staining correction factor of 0.1 as determined previously (19 (link)), which we use to correct all gel data.