1. To increase resolution of the DNA bands:
a. Running the gel at a lower voltage for longer period of time
b. Using a wider gel comb
c. Loading less DNA into well
2. To get better separation of bands, adjust the agarose percentage of the gel if the similarly sized bands that bands that are running too close together are loaded. A higher percentage agarose
gel will help resolving smaller bands from each other, and a lower percentage gel will help separating larger bands.
Protocol for 1% Agarose Gel
A. Pre-cast Protocol (same like as EtBr)
Prepare agarose gel solution using your standard protocol.
To prepare the gel, add 5 μL of Gel RedSafe nucleic acid dye (10,000 x, water soluble) per 50 mL of agarose gel and mix thoroughly.
Gel RedSafe has excellent thermal stability and can be added directly to the high-temperature gel solution without waiting for the gel solution to cool.
As an alternative: by pre-mixing Gel RedSafe with an electrophoresis buffer containing agarose powder and heating uo.
Note: The pre-cast protocol is not recommended for polyacrylamide gels. Use the post staining protocol for acrylamide gels.
B. Post-staining Protocol
1. Electrophoresis according to conventional methods.
2. Dilute Gel RedSafe Nucleic Acid Dye 10,000 x stock solution approximately 3,300 times to make a 3 x staining solution (for example, add 15 μL of Gel RedSafe Nucleic Acid Dye 10,000 x stock
solution to 50 mL of H2O).
3. Carefully place the gel into a suitable container and soak the gel with a sufficient amount of 3 x staining solution. In order to shorten the soaking time, the dyeing solution can be
pre-heated to about 70℃, then put into the gel and incubate for 10 min to obtain the ideal effect.
(if not heated, incubate for 30 min in room temperature shaker; if it is acrylamide gel, incubate for 30-60 min and extend with the increase of acrylamide content).
The amount of bubble dyeing dye is large, and the dyeing solution can be reused for about 3 times for a single use. The 3×Gel RedSafe stain solution can be prepared in large quantities and stored
at room temperature until used up.