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Library was constructed using Clontechs SMART cDNA library construction kit. RNA was DNase treated prior to 1st strand cDNA synthesis. 0.5ug of total RNA from pooled 35-day dorsolateral prostate, 0.5ug of total RNA from pooled 35-day ventral prostate, and 0.5ug of total RNA from pooled 35-day coagulating gland (anterior prostate) was combined and used in 1st strand cDNA synthesis. 2nd strand cDNA was generated by 21 cycles of PCR, digested with SfiI and ligated into the lambdaTriplEx2 phagemid. Phagemids were converted to pTriplEx2 plasmids in the BM25.8 e-coli strain. Created from pooled prostate from five 35 day old male C57BL6 mice by Colin Pritchard.
Nucleotide
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