AMBIONS ANTISENSE 18S TEMPLATE

K.Kramer 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 
> titrate)
> 
> 
> 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 
> once.  
> 
> 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.
Kerstin>
> > > 
> 




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