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PCR alternative to plasmid rescue?

gvacano at eagle.wesleyan.edu gvacano at eagle.wesleyan.edu
Mon Feb 21 11:41:21 EST 1994


Hello--

	Our lab has been doing P-element transformation for some time now,
using pCaSpeR vectors (esp. 3 and 4). We've obtained a few transformants that
are homozygous lethal, and we'd like to obtain sequence information for the 
region(s) in which the P-element cassette(s) are located. Since vectors like 
pCaSpeR are not conducive to plasmid rescue, apparently some workers have 
utilized a PCR technique, involving digestion of genomic DNA, circularization 
of the resulting fragments, and amplification using cassette-specific primers, 
which point away from each other, to amplify genomic DNA flanking the 
P-element cassette.

	I would like to do the same, and I was wondering if anyone could help
me. Specifically, I need pCaSpeR sequence information, information concerning
pCaSpeR specific primers (primers that recognize the P-element inverted repeats
would be extremely helpful), and any protocols, hints, tips, advice,
references, etc.

Thank you, Guido



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