RFI: solution DNA hybridization?

Don Chen chend at ucs.orst.edu
Wed Nov 26 01:35:59 EST 1997


Hi:

We would like to probe some transgenic plant DNA for incorporation of
transforming DNA.  While I've done solid phase Southerns, I don't know
of a protocol for binding probe DNA to genomic DNA in solution.  Is
there one out there?

Assuming there is one out there, we would like to use biotinylated
dNTP with a random priming kit.  This would be hybridized in a
microfuge tube with the genomic DNA.  The hybridization would be
scored yes/no by binding the biotin to a strepavidin coated ELISA
plate.  We would follow up with SA-alkaline phosphotase and then
substrate.  The scoring would be done on an ELISA reader.  

Does this sound feasible/reasonable?

Thanks in advance for any help.

Don

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Don Chen            *  Standard disclaimers apply here.
USDA-ARS-NFSPRC     *
3450 SW Campus Way  *   
Corvallis, OR 97331 *
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