UV-Cross

Kerstin Reisinger Reisinger at em.uni-frankfurt.de
Mon Feb 14 06:47:25 EST 2000


Hey folks,

we`re trying to do UV-Crosslinking at the moment, but only trying
because beside the bands we have a lot of smearin the lanes.
We´re using a Stratagen Stratalinker 2400. The DNA oligo is 28 bp and
labeled with klenow.
To get rid of the background we changed the energy and the distance
between the probes and the `lamps. We used pdI-dC or salmon sperm DNA in
variable concentrations. The proteinamount of 20 microgramm seemed to be
O.K.
The running gel has 10% polyacrylamid and normal Laemmli and sample
buffers are used. Before drying the gel is fixed in a mixture of acetic
acid, methanol and H2O.

Does anyone used this technique and have some good ideas for us to
overcome this problem?

Kerstin





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