PCR KIT-> NO GELLS, Quantitative, Double Fidelity

gwade at physiology.uwo.ca gwade at physiology.uwo.ca
Thu Dec 7 17:30:11 EST 1995


odonnell at sasa.gov.uk wrote:


>> Attention PCR USERS
>> 
>> I just found a great new product from Kalyx Biosciences.  AMPLISCRIPT 
>> 
>> allows you to detect your PCR products using an ELISA format.  

>This sounds interesting.  Can you give a few more details about it? Does it involve specially labelled 
>primers for example, or can plain PCR primers be used? Does it work 
>on 96-well plates?

>Kevin

>Kevin O'Donnell
>SASA
>Edinburgh

No labelled primers.

Works on 96 welled plates.

Its actually quite a simple process involving using a T7 promoter
region on your probe (primer).  After pcr one will have generated
multiple copies of the intended gene.  The user then makes
complementary RNA copies of this code (thanks to the T7 promoter now
being on the amplified DNA from the original PCR).

So now you have many copies of RNA which is complementary to the DNA
you were probing for.  Perform a nucleic acid hybridization with some
control DNA and you get heteronucleic acid (hNA).  Kalyx has
antibodies to the hNA allowing one to determine the presence of this
molecule.  

All the reagents required to do this (except TAQ) are provided.  I
hope the way I have explained it is possible to follow.  

Kalyx has some literature outlining this process.

Interestingly enough they also have an inexpensive matrix on which you
can do a nucleic acid hybridization in about 90 min (at room temp).

good luck

GREG WADE
Physiology PH.D Candidate
The University of Western Ontario
London, Ontario Canada.






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