Need help for in-vitro DNaseI-Footprint

Fritz Schwarzmann Fritz.Schwarzmann at gmx.de
Wed Aug 25 08:31:31 EST 1999


Hi all!

We want to establish DNaseI footprint experiments in our lab.

We already see very weak protection by bound proteins, however, the
footprints are heavily masked by strong background signals.
This background is also visible in the control lane with the labelled probe
(approx. 500bp long) prior to incubation with DNaseI. The probe has been
freshly prepared and labelled (terminal transferase or Klenow). The
solutions were also checked for DNase contamination and freshly prepared
several times.

Did anybody else see such kind of degradation of the labelled probe? How can
this be prevented?

Thank you in advance.

Fritz

University of Regensburg
Germany





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