RediVue and EasyTides..Good?

tflanigan at molbiol.ox.ac.uk tflanigan at molbiol.ox.ac.uk
Mon Nov 13 07:10:10 EST 1995


In article <47tlme$48j at reeve.research.aa.wl.com>, Angela Hofstra <Hofstra at aa.wl.com> writes:
> paquette at shaman.nexagen.com (Tom Paquette) wrote:
>>We are looking at using room temperature 32P-gamma-ATP in an Amersham
>>dispenser.  We use several millicuries per week but have had a couple
>>of instances where the free PO4 as assessed by PEI sheet is 40 or 50%.
>>It is clear that precise use conditions may be of relevance here, perhaps
>>utmost relevance. E.g. the dispenser only does 10ul per shot and someone
>>needing 20 ul might carry PNK back into the storage ampule.  Now I never
>>heard of anyone mixing kinase or any other reactions in an order other
>>than A-B-C-32P-enzyme and we did put up a sign that says you have to pipet
>>32P into clean tubes. But the question is whether anyone else has had
>>good or bad experiences with these room temperature 32P nucleotides from
>>NEN and Amersham?  On the surface of it, and especially for us, they seem to
>>be significant advantage since they are quick and easy to use and ought to
>>be replaced on a weekly basis.  Thanks
>>
>>Tom Paquette, PhD
>>Senior Scientist
>>RSO
>>paquette at shaman.nexagen.com
>>NeXstar Pharmaceuticals
>>Boulder, CO
>>303-546-7626
>>
>>
> 
> Tom,
> 
> We use redivue P32 from Amersham regularly.  We don't use a dispenser as 
> you describe it. Anyway, I think the convenience is great and we get good 
> labelling.  The pink colour helps to keep track of where your label is.  
> I've even used it a week after delivery (but only3 days after the 
> calibration date) and still got good Northerns.  
> 
> Best of luck,
> 
> Angela
> 
Tom,
we've looked at using redivue and easytides in differential display of mRNA
and they did not work.  The preservative(?) seems to interfere with the PCR 
reaction.  We were also told that these nucleotides did not work well for in 
situ hybridisation but we did not try it.
Cheers,
Tom Flanigan



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