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gel shift

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SAM

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Sep 5, 1996, 3:00:00 AM9/5/96
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I'm having problems with the competition assay on the gel shift i.e. the
cold oligo does not block or even decrease the binding of the hot one
when incubated with a rat liver nuclear protein extract. However, the
bands are clear and nice and on the same level as the HeLa cell control
(competition assay works for the comercial HeLa extract). Did anyone
have a similar problem and/or has any ideas on what could cause it?
Thank you.
Anna Liachenko
Mcgill University, Montreal, Canada

martin LEACH

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Sep 6, 1996, 3:00:00 AM9/6/96
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Could be tons of nonspecific binding....

try performing a poly dI-dC titer or add another nonspecific DNA such as
sonicated salmon/herring sperm DNA....to see if they cause these bands to
disappear...

just my thruppeny bit

M

SAM (b7...@musicb.mcgill.ca) wrote:
: I'm having problems with the competition assay on the gel shift i.e. the

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