I have recently shifted from EtBr to SYBR GOLD for staining my DNA in agarose gels. But the 1kb+ DNA ladder in the SYBR GOLD gels are melting, rendering them useless. I am assuming this has something to do with the SYBR GOLD stain which I add to the agarose mixture before I cast then gel.
I would appreciate any suggestions/solutions
I have noticed that out 1kb+ ladder runs horribly in gels stained with SAFE VIEW. I have switched to Generuler by Thermoscientific and this works well. So in answer, I don't know why but we seemed to have resolved the issue!
We always stain our gels with SybrGold AFTER the run. This way there is no interference, staining is evenly distributed and the sensitivity is amazing. If sensitivity is not really an issue then I recommend SybrSafe which doesn't seem to interfere with the DNA.
Yes..I agree with Jesper. Stain with SybrGold after the run. It is highly sensitive allowing you to visualize even lower amounts of DNA on gels.
I was having this problem, and staining the gel afterward fixed it right up. I found a good resource which touches upon why. Basically the SYBR Gold retards the movement of DNA through the agarose. https://tools.thermofisher.com/content/sfs/manuals/td004.pdf
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