PCR DIG-labeling Southern probes

Della A Johnston dajohnst at uoguelph.ca
Thu Aug 7 06:57:59 EST 1997


I would suggest using the whole plasmid. You will get a stronger signal 
since more label will be incorporated.

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	Della Johnston
	Dept. of Pathobiology, Ontario Veterinary College
	University of Guelph, Guelph, Ontario  N1G 2W1
	Telephone:  (519) 824-4120   ext. 4630
	Fax:  (519) 824-5930
	email:  dajohnst at uoguelph.ca
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SVLUsagi2 (svlusagi2 at aol.com) wrote:
: I have a 514-bp insert in a pUC12, 2,680-bp, plasmid that I use as a DNA
: probe for a Southern blot against rabbit genomic DNA.  I have started
: using Boehringer-Mannheim's PCR DIG-labeling synthesis kit for
: digoxigenin-labeling of this probe.  Is it better for me to cut out the
: 514-bp insert from the plasmid 
: (using Pst I), gel purify it, then use this as the DNA template for the
: PCR DIG-labeling reaction, or can I just use the whole plasmid as the DNA
: template?  Thanks in advance for any suggestions.



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