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270ug total RNA was extracted from 200 adult male mouse retinas. A directionally cloned cDNA library in the pSPORT1 vector(Life Technologies) was constructed at Bioserve Biotechnology (Laurel MD) essentially following the protocols of the SuperScript Plasmid System full details of which are contained in the manufacturer's Instruction manual (http://www.lifetech.com/). First strand synthesis was carried out using a Not I primer-adapter [5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCC(T)15-3']. cDNA was cloned in Not I/Sal I sites. EST analysis was performed on the unamplified library at the NIH Intramural Sequencing Center (NISC).
Nucleotide
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