Footprinting

Greg g.dean at uea.ac.uk
Fri Jul 3 12:08:22 EST 1998


Hi All,

I'm trying to footprint several regions of  my favorite promoter, and am
using the Pharmacia kit. I am attempting to bypass the shift step and go
straight to the footprint. I have one problem I can find little in the
literature to give me a rough idea of how much protein I should be using
in an individual reaction; I am going to opt for approx. 25ug per rxn of
a crude nuclear extract, as this fits with previous shift work done in
my lab, with the felling that an a smallish excess is far more
preferable than being low on protein. I have to titrate my DNase I yet,
my job for Monday morning. So any suggestions about this or any little
help about footprinting in general would be greatly received as I have
no one with experience of this technique around me.

Thanks in advance

Greg.
----------------------------
Dr. Greg Dean
Dept. of Biological Sciences
University of East Anglia
Norwich
Norfolk
NR4 7TJ England
Tel.: 01603 593796
Fax.: 01603 592250
----------------------------





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