DIG oligo 5'-end labeling

Richard T. Pon rtpon at acs.ucalgary.ca
Tue May 2 13:03:56 EST 1995


ag296 at FreeNet.Carleton.CA (Tracey Forster) wrote:
>
>I currently use 32-P to end label my oligo rRNA probes
>(17-21-mers). Because I required quantification this was the
>easy and obvious route.  I now have a need to use these probes
>for routine detection from colony lifts and I was considering
>using DIG labeling.  BMC has a kit for end labeling
>amino-substituted oligos using digoxigen-NHS-ester, has anyone
>tried it?  Does it give acceptable results? any drawbacks?
>
>Bob Forster
>Centre for Food and Animal Research
>Ag Canada
>Ottawa, ON

Dear Bob
	The DIG-NHS ester reagent sold by B-M works well for
end-labeling 5'-amino oligonucleotides. In fact, we use this
reagent as part of our routine DIG-labeling service
($100 CDN plus basic synthesis costs, price includes
purification by gel electrophoresis). The labeling reaction is
just a matter of combining the oligo and DIG-NHS reagent together
(in DMSO/water) and letting sit overnight. We separate unreacted oligo
and excess DIG from the labeled product by polyacrylamide gel
electrophoresis, but it could also be done by HPLC.
	If you want more information about our synthetic services,
send me your mailing address or fax number.

Richard T. Pon
Director
UCDNA Services, The University of Calgary
tel: (403) 220-4277, FAX: (403) 283-4907



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