plasmid integrated or not ?

Jarmo Niemi jarnie at utu.fi
Tue Mar 15 03:32:51 EST 1994


In <2m30lr$mi3 at agate.berkeley.edu> AT (Theo at mendel.Berkeley.EDU) wrote:
> The following question was posted to me by a friend. I told

> How can you test if the plasmid vector is integrated into
> the genome of transfected cells ? (Only 5% of cells are
> transfected and there's no selection marker on the vector).

Prepare genomic DNA from suspect clones, digest with a couple 
of restriction enzymes, which cut the plasmid few times, run a
gel with the plasmid cut with the same enzymes as controls, do
a Southern hybridization with the plasmid as probe. You can 
probably even locate the position, where the integration has
taken place.

Jarmo Niemi   Biochemistry, University of Turku
              SF-20500 Turku, Finland
              ++358-21-6336856
	      FAX ++358-21-633 6860
              jarnie at utu.fi

	Also janiemi at finabo.abo.fi



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