Question about gel shift assay
mfm434 at rzmail.uni-erlangen.de
Thu Jun 20 12:29:23 EST 1996
Philippe Lefebvre wrote:
> Using shorter probes will help but in our experience, this will not allow you to get
> rid of non spec binding. You 'd better try to use different non spec DNA such as
> salmon sperm DNA, calf thymus or poly dIdC. Results can be really strikingly different! ^^^^^^^^^^
Poly dIdC is absolutely necessary to get rid of non spec binding....
I have accidently done it once without: => terrible!!!!!
Use 2 micrograms max (more does'nt make any further effect!)
Probe lenght: i use normaly very short probes; up to 25 b for a single transcription factor
If you want a complete protocol that works fine, pls E-mail me...
Dipl. Biol. Heinz-Juergen Schaefers
Universitaet Erlangen-Nuernberg / Med. Klinik IV / Nephrologisches Forschungslabor
Loschgestr. 8, D-91054 Erlangen, Germany
E-mail: dienstl.: mfm434 at rzmail.uni-erlangen.de / priv.: h-j.schaefers at erlangen.netsurf.de
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