SYPRO orange stain on PhastSystem gels ?

Vadim Kraynov vkraynov at pop.service.ohio-state.edu
Wed Jul 9 21:33:09 EST 1997


Well,

I tried to stain another Phast System gel with SYPRO today.
Stained for about 30-40 min; and then sliced the gel off the plastic backing 
with a razor blade.  Upon transilluminating with UV, it does fluoresce and 
the bands are visible; however, the sensitivity was definitely lower than the 
silver (I ran two identical gels, and stained one with silver, as usual, in 
the PhastSystem).  So it does work, and for quick staining, it probably is a 
viable alternative.  But with sensitivity close to Coomassie, ability to make 
a quick photo is it's only advantage. 

Also, until I find a better way to take the gel off the backing (it breaks 
easily upon slicing off), it's pretty inconvenient.  I guess, I'll need some 
sort of small "cheese slicer" or some such thing.

By the way, illumitating with UV from above doesn't really help to visualize 
the bands, at least, not with the lamp we have.


>:Thanks for this suggestion ! You are probably right - I 'm not sure what the 
>:gel backing consists of, but it probably does block UV.  I was thinking about 
>:removing the backing somehow, but the gel is really fragile, and I couldn't 
>:find the way.  I'll try illuminating it from above.
>:
>:The problem , however, is that I NEED to TRANSilluminate in order to 
>:photograph.
>:
>: This dye, if works, would provide a great alternative to time-consuming 
>:silver staining, with  practically the same sensitivity, plus easy 
>:"photographability" - just like regular EB stained agarose DNA gels.   Cheers
>:
>:Vadim
>
>Lured by the same prospect, I tried the dye ~ 2 years ago (from Molecular 
>Probes). In the same gel imaging system as used for DNA gels, the staining 
>was only ~ as sensitive as colloidal coomasie. To get it to be as 
>sensitive as silver one needs (Ok, in my experience at least) to fiddle 
>with different filters/UV sourses - something I didn't like and never 
>will. My conclusion is that it does not work as advertized. 



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