Good afternoon,
Propidium iodide (PI) is a dye use to stain nucleic acid in flow cytometric analyses, particularly in cell cycle analyses. Has anyone dared to try staining a DNA electrophoresis gel this dye (PI)? Would it be worth trying?
Thanks in advance
I'm sure it will work, but you will have to use different wavelengths for excitation and emission than you would use for ethidium bromide. Thermo-Fisher gives the wavelengths for DNA-bound propidium iodide as 535 and 617 nm.
Dear Luis
as Adam told you in principle it will work because ot is able to bind dna and it is fluorescent, and if i remenber well it was used in come assay approach which is based on agarose gel but using a fluorescence or confocal microscopy whih allow you to select the optimal waveleghnt for it.
For routine agarose gel analisys, if it work well or not depends from the source that you would like to use for visualize the gel and from the emission that it can have when excite from this source.
for example it can be efficiently excite with UV trasnilluminattor (excitation beetween 250 and 350nm) or with blue light ( about 450-470nm)
from the attached paper seems that fire Red staining is based on propidium iodide https://www.abmgood.com/Safe-DNA-Fire-Red-Stains.html and it seems to work well.
However personally as alternative to ethidium bromide i like a lot the Gel Red of Biotium ( sole also by vwr) which is quite cheap, less toxic and very stable under uv excitation and with exposition to enviroment light.
ciao
Manuele
Dear Manuele,
It was in fact reading Safe Fire Red info what made me wonder about PI use in gel electrophoresis. It was pure curiosity with the intention of having an alternative (ideally cheaper) for DNA staining. However, given the points mentioned by you and prof. Shapiro along with some info I got by myself, I won't take my curiosity to practice this time. Guess I'll go with Gel Red afterall.
Thank you very much
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