We have always used EtBr in our lab, but due to changes in institutional policies, we are migrating to GelRed.
We are aware of the possibilities of Post-staining protocol and Pre-cast protocol of GelRed, but we want to rule out all possibilities and test also the direct application of GelRed in the samples.
What would be a good proportion of GelRed/Samples volume?
Thank you very much!
According to Uribe et al. (2013) Preparation of the GelRed reagent and visualization of DNA samples: For the preparation, 3 µl of GelRed (GelRedTM Nucleic acid gel stain, 10,000X; biotium) a 997ul of loading buffer (60% glycerol, 0.05% of bromophenol blue) obtaining a final concentration of 30X according to the manufacturer's recommendations. Once the loading buffer was prepared with the GelRed reagent, Two 1% (w/v) agarose gels were prepared. stained with ethidium bromide (5ng/ul)5 and with him GelRedTM reagent respectively, the latter was added to each total DNA sample and to each PCR product in a 1:1 ratio v/v (GelRed loading buffer: DNA); Finally, The gels were visualized in the High Performance ultraviolet light transilluminator. Ultraviolet transilluminator (UVP).
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