Help with UV Shadowing requested
drm21 at mole.bio.cam.ac.uk
Thu Oct 26 09:53:54 EST 1995
In article <46lt28$n57 at taco.cc.ncsu.edu>, smlane at unity.ncsu.edu (Stephen
Marcus Lane) wrote:
>Hi, I'm a grad student in Biochemical Engineering here
>at NCSU, and I'm having real trouble getting a
>UV shadowing protocol to work. I haven't been able
>to find much at all in the literature or protocol
>manuals on the technique.
>UV shadowing is supposed to work by running RNA or SS DNA
>on an acrylamide/urea gel, then exposing the saran-wrapped
>gel to UV light. The light is absorbed and fluoresced
>by a TLC plate (I'm using Brinkmann: Merck silica gel
>F254), except where the RNA/DNA is on the gel. In these
>spots the RNA/DNA absorbs the flouresced light, supposedly
>showing up as "shadows" on a green background.
One other thing (than the only other reply I've seen...) -
Are you getting the geometry of your set up right?! It needs to go:
GEl on saran wrap
TLC plate (or other fluor)
ie, your gel must be between the UV source and the fluor - the RNA/DNA is
NOT absorbing the fluoresced light, it is absorbing the UV and preventing
it exciting the fluor. Hope this helps,
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