Determining Plant Transgene Copy Number

Kyle Anderson kyleande at gpu.srv.ualberta.ca
Mon Aug 23 18:22:49 EST 1999


I work in a canola lab and our T0 generation are starting to die out before
we can determine which plants have a single copy of our gene.  We are
currently doing genomic extractions, cutting with EcoRI and then Southerns
to determine copy number.  I was wondering if anyone had ideas about the
possibility of cutting the genomic DNA, diluting and recircularizing, and
then running PCR using primers which extend away from our gene.  It would
seem to me that this would give a different band for each insert, but would
this work with genomic DNA?  Is it feasible to try and circularize all the
digested products of genomic DNA?  Any comments you may have about
determining copy number in plants would be greatly appreciated.

Kyle Anderson
kyleande at ualberta.ca
geneng at arc.ab.ca








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