Hot cDNA Probes???

rothman at mcclb0.med.nyu.edu rothman at mcclb0.med.nyu.edu
Wed May 11 17:36:20 EST 1994


Hello to all!!!

We here in the Big Apple are curious about using reverse transcribed
cDNA as a probe to screen a dot blot.  We have several kinase genes we
now know to be expressed by different classes of pediatric brain
tumors.  We are now ready to screen a larger pool of tumors for these
genes (mostly receptors).  

Our thought was to dot-blot cloned DNA, then reverse transcribe cDNA
from total tumor RNA (in the presence of 32P) and use this as the
probe against the blot. 

Does that sound even remotely feasible?  We worry about
quantity...tumor tissue, and hence RNA, is very rare and usually we
only get small chunks.  Any advice or protocols regarding this "hot"
cDNA idea would be greatly appreciated.

Thanks,
	Marc Rothman
	Howard Weiner
	Howard Hughes Med. Inst.
	Dept. of Biochemistry
	NYU Med. Center
	550 First Ave.
	NY, NY  10016
	212-243-6753
	212-683-8453 (fax)



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