[Drosophila] technique suddenly not working

Kathleen Gajewski sa08383 at wotan.mdacc.tmc.edu
Fri Nov 11 17:01:39 EST 2005


I'm suddenly having trouble with the Gloor and Engels technique for PCR 
from single fly genomic preps (I'm trying to work out a PCR test for 
verifying transgenic lines).  I've remade my solutions- no luck.  I've 
included fly strains/primer combinations/PCR conditions that have 
worked in the past-still no luck.  I've included positive controls of 
the purified construct DNA, and those work just fine, so I know the PCR 
itself is not at fault.  Does anyone have some trouble shooting 
advice??  Usually the technique works just fine,  but every now and 
then it just fails out of the blue and I'm at my wits end with this!!

Thanks

Kathleen



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