AMBIONS ANTISENSE 18S TEMPLATE
ugha083 at alpha.qmw.ac.uk
Thu Aug 22 06:59:21 EST 1996
On 19 Aug 1996, Jennifer L Causey wrote:
> I have been having trouble with the T7 18s transcribed probes from Ambion
> on RPAs. I can not seem to:
> (a) get the correct specific activity so it will show up but not so much
> so it wont overexpose.
> (b) transcribe enough probe to saturate the RNA sample (it does not
> p.s. my other probes are really low expressed so I have to expose for 3
> to 7 days to see them and I want to do densitometry on them and 18s at
> any suggestions for the amount of hot/cold labeled nucleotides to add?
> any experience with this peticular template would be helpful.
> also anything to do with porcine control probes that have worked for you.
> please respond.
> - L.
I had similar experiences with this template, try to religate and
and do a transfection, that would also save you bying it again. As for
the labelling it depends on the abundance of your message, but something
in the range of a 1 in 100-300 dilution of your radioactivity should do.
> > >
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