I am in the process of conducting an electrophoresis and I have a Safer dye at my disposal. However, I am uncertain whether it would be more suitable to combine the Safer dye with the 6X DNA LOADING Buffer. Would it be advisable to include the Safer dye in the loading DNA buffer mixture before incorporating it into the DNA sample? Or is such a procedure unnecessary?
I assume you mean it's a stain that is a safer alternative to Ethidium bromide, like SYBR Safe?
You do not need to mix it into your DNA loading buffer or the DNA samples. Just add the SYBR to the melted agarose, swirl well to mix, and pour the gel.
Hi,
You can add SYBR Safe (or similar) to either the gel as suggested above, or to your electrophoresis running buffer. You can even incubate with the dye for ~10 min after your run is complete.
A word of caution, it's called 'SYBR safe' but actually only marginally safer. The gel and buffer should be taken care similarly to an EtBr gel and go to biohazards.
Hi there,
the more cautious protocol is to run the gel without any intercalating agent (so first your apparatus and fluid remain safe and second the mobility of DNA is not affected by intercalation) and then to put your gel in running buffer containing the intercalating dye. As already mention any so-called "safe" intercalating agent is not safe at all and therefore vessels, gels and fluids should be treated accordingly.
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