Gel Shift Probe

Steve Rodems smrodems at students.wisc.edu
Fri Apr 1 01:01:51 EST 1994


Iv'e been seeing some strange things with 32P labeled probes in my gel
shift experiments.  In a control lane with no extract added my probe is
consistently two bands about 1 cm apart at the bottom of a 5-10%
polyacrylamide gel (all other lanes have the same thing).  Sometimes I get
a couple other bands above and below the major ones.  I fill in 5'
overhangs (4 nt) to label, my label is the first nucleotide added in the
fill in, and it labels only once on each end.  The DNA I use has been
either gel purified 30 bp or two 30 nt oligos annealed.  I thought that
maybe it could be incomplete fill in so that I have a range of labeled
probes different by 1-4 bps but I wasn't sure if the separation would be
that great.  Does anyone have any ideas or has anyone seen this before?


-- 
Steve "Some day I will get the hell out of Wisconsin" Rodems

"Then I am here for the Lee family renioun ... shur-wajo-shur"



More information about the Methods mailing list