Non isotope Northerns

Michael Sullivan via methods%40net.bio.net (by mlsulliv from wisc.edu)
Thu Jun 21 14:06:27 EST 2007


I'm not sure if there have been any improvements in the non- 
radioactive detection systems in the past few years, but about seven   
years ago I did a side-by-side comparison of DIG-labeled probe  
(ultimately detected with an anti-DIG antibody fragment and a  
chemiluminescent detection system) with standard isotope labeled  
probe. I did the comparison at the urging of the sales rep for the  
company selling the non-radioactive system.

What was most disappointing about the non-radioactive system was that  
it was quantitative over only a very narrow range (around 3 fold I  
recall) compared to the several orders of magnitude linear range of  
quantifying radiolabeled probe using a storage phosphor  screen. For  
me this was not acceptable

Have you looked into other options such as real time qPCR?

Mike Sullivan


On Jun 21, 2007, at 11:46 AM, hilary cassidy wrote:

> Hello!
> I was wondering if anyone out there had any experience of doing non
> isotope northerns? Can anyone comment on the sensitivity of method
> compared to the traditional isotope methods or what is the threshold
> concentration you need to be able to detect anything? I am trying to
> persuade my boss to let me try this instead of the isotope methods as
> I am returning to my home college where they don't have an isotope
> licence. Any help or advice would be great!
>
>
> Cheers
>
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---
Michael L. Sullivan
Plant Research Molecular Geneticist
US Dairy Forage Research Center
ARS-USDA
1925 Linden Drive West
Madison, WI 53706
(608) 890-0046 (Phone)
(608) 890-0076 (FAX)



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