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1st strand cDNA was primed with oligo(dT)17 on 50 ng of DNAse-treated, total cellular RNA obtained from 5,000-10,000 microdissected cells histologically-determined to be fully malignant prostate cancer cells. Double-stranded cDNA was ligated to EcoRI adaptors, 5 cycles of PCR applied to the cDNA with an adaptor-specific primer, and the resulting PCR product subcloned into pAMP10 by the UDG-cloning method (Life Technologies). Average insert size is 600 bp. NOTE: Not directionally cloned. This library was constructed by David Krizman.
Nucleotide
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