HELP: Problems probing for T-DNA in Southern Blotting

Nancy Fineberg fineberg at ucalgary.ca
Wed May 23 09:58:04 EST 2001


Hi All,

I am having difficulty detecting either my transgene or the selectable
marker within the T-DNA inserted into a plant genome, both are synthetic
genes.  We know from protein purification as well as PCRing & sequencing
the gene  that it is there!  Also, I have done Southern blots on a
single copy native dehydrogenase, and had no trouble with detection.  I
am using the High Prime kit for labelling and the QuickHyb buffer for
hybridization.

Any advice would be greatly appreciated.
Nancy Fineberg
Biological Sciences
University of Calgary
fineberg at ucalgary.ca

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