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EZ-Vision vs Ethidium Bromide: Comparison of Fluorescent D yes for the Visualization of DNA Bands in Agarose Gel. Experimental Method. Used plasmid DNA, [ pDNA ] = 0.2ug/ uL Made two 1% Agarose Gel: 1 with EtBr & 1 without EtBr
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EZ-Vision vsEthidium Bromide: Comparison of Fluorescent Dyes for the Visualization of DNA Bands in Agarose Gel
Experimental Method • Used plasmid DNA, [pDNA] = 0.2ug/uL • Made two 1% Agarose Gel: 1 with EtBr& 1 without EtBr • Prepared 6 samples of pDNA to be loaded in each gel with different amounts of pDNA : 0.01ug, 0.05ug, 0.1ug, 0.5ug, 1.0ug & 5.0ug • In gel without EtBr, added 1uL of EZ-Vision to the 6 samples and the 1kb DNA ruler before loading • Ran gels at 120V for 1.5-2 hours • Imaged them
Results EtBr Gel EZ-Vision Gel 1 kb DNA ruler 1 kb DNA ruler 0.05ug 0.05ug 0.01ug 0.01ug 1.0ug 1.0ug 0.1ug 5.0ug 0.1ug 5.0ug 0.5ug 0.5ug
Conclusion • EZ-Vision fluorescent dye can be used as a safer alternative to visualize DNA bands in an agarose gel • EtBr is more sensitive at detecting DNA at lower concentrations (<0.1ug) when compared to EZ-Vision • EZ-Vision gel could be used on the UV transilluminator to cut bands – tested and found that you can detect the bands just as clearly as if they were stained with EtBr.
How to add EZ-Vision Dye • Prepare samples and DNA ruler to be loaded on gel in usual method. Add 1uL of EZ-Vision dye to each before loading. • Mix sample to get homogenous solution. • Load and run gel. • Image gel. Remember to switch emission filter to SybrGreen before using imaging program. • Capture photo, save and/or print. DONE!
