Why does in situ work with sense probes, too?
MCOLLINSON at lab0.vet.ed.ac.uk
Tue Dec 17 03:44:08 EST 1996
Julian Dow <gbaa02 at udcf.gla.ac.uk> wrote:
> My wife asked me to post and ask the big unanswered question about in
> situs: why is is a generally acknowledged (but not widely published
> ;->) fact that for many genes, both sense and antisense probes give the
> SAME, tissue-speicific, staining patterns?
> Are there reverse transcripts, or what?
Whilst reverse transcripts could, of course, occur, it is more likely
that something has gone wrong during transcription of the probe. Maybe
one possibility is that if you have cut the plasmid with an enzyme
that gives a three-prime overhang, you could get RNApol binding leading
to transcription of the opposite strand of the plasmid i.e. the
sense control could contain appreciable amounts of antisense RNA.
I'm sure other people have got better ideas, though.
Martin Collinson (MCOLLINSON at lab0.vet.ed.ac.uk)
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