T7 transcribed RNA probes

Torsten Boerchers borcher at uni-muenster.de
Sun Jan 14 08:45:46 EST 1996

Hello Northern or in situ hybridization specialists

Anybody else using T7 RNA polymerase to transcribe
RNA from a DNA template (e.g. linearized plasmid with T7-
promoter) who also observes multiple bands in a gel or blot?

We use this technique for generation of DIG labelled 
RNA probes and would like to know whether some 
people have explanations for this behaviour or 
are aware of published evidence.

In some manuals it is emphasized not to allow 3´overhangs
in the restriction made for linearization. The overhang 
could be an initiation point for the T7 RNA polymerase. 
Any mechanism for this known?

Thank you for any help

                          Torsten Boerchers                            /
    _/  _/_/_/ _/_/_/    Institute of Chemical- and Biochemical Sensor/
   _/  _/     _/   _/   Research (Molecular Biology Group)           /
  _/  _/     _/_/_/    Mendelstr 7, D-48149 Muenster, Germany       /
 _/  _/     _/    _/  Fax: +49-251-980 2890 Phone: +49-251-980 2880/
_/  _/_/_/ _/_/_/_/  E-Mail: borcher at uni-muenster.de              /

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