I am struggling with getting quality results from my COMET assays, and have been using DAPI as a stain. Is there any reason DAPI can't be used? I'd rather not use EtBr but will if DAPI is the issue.
Sure, just use some antifade medium for the reduction of the photobleaching if you want to gain comparable quantitative results!
After electrophorsis wash several times with deionized ultrapure water then Dehydrate gels with absolute ethanol , let it dry then stain with Etidium Bromide 10 micrograms per ml whatever you want for 5 min before visualize it and to capture images at 10X.
Dear Brenna Hay,
You can use any DNA dye.
However, you should consider that different dyes have different detection limit.
If you are interested in very precise analysis it is better to use SYBR® Green or Vista Green®. But, DAPI is still suitable, be careful with DAPI Excitation⁄Emission wavelength 358⁄461 (nm)
dehydrated gels and stained eg. propidium iodide 10 microg per ml for 5 min in dark , allows you to image capture several areas at 10X with several comets almost in the same focus plane very fast so photobleaching of the dye should not be a problem. You do not need to use an expensive budget.
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