re; non-radioactive DNA detection

37_410 at uwovax.uwo.ca 37_410 at uwovax.uwo.ca
Tue Jul 24 10:28:38 EST 1990


J. Sternberg writes:
>Subject: non-radioactive DNA detection
>From: J.STERNBERG at aberdeen.ac.uk
>Date: 18 Jul 90 11:04:09 GMT
>
>I hope to put a student, who won't have clearance for P32
>work for some while, onto doing some restriction mapping
>of the ribosomal genes in digenean parasites shortly. So it
>seems like we will have to use non-radioactive labelling
>techniques on our probes, and I have no experience in
>this area. I have a nice glossy booklet from
>Boehringer describing their DIG system. It looks OK
>but then it should be at the price. I wonder if any
>of you have any experience with this or other
>non-radioactive labelling methods and any comments
>on their sensitivity.
>If I get enough responses, I'll collate them and mail
>them back to the bboard,
>Thanks, Jeremy Sternberg <J.Sternberg at uk.ac.aberdeen>
>               <J.Sternberg%uk.ac.aberdeen at ukacrl> (


	The limited use that we have had with this DIG system relates
to its use in undergraduate labs to do Southern blotting of bacterial
genomic DNA although we do plan to use it in our own research in the 
near future.  We have had no problems with the system and in fact I 
have changed many of the parameters around with no apparent problems.
The changes I made were so that I could use the kit in conjunction 
with the Hoeffer "Hybrid-eze" system which makes use of different 
volumes than those given in the DIG kit instructions.  Furthermore, I
have coupled the kit with the use of the LKB/Pharmacia "Vacugene" 
system such that less time is required overall from actual blotting
to hybridization.  As I have had no problems I feel that I can highly 
recommend the product.  The results have always been clean, quick 
and reproducible.

			Lou Passador
			c/o T.LINN at uwovax.uwo.ca
			U. Western Ontario
			London, Ontario
			Canada.



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