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Approximately 1mg total RNA was extracted from 200 adult mouse whole eyes. A directionally cloned cDNA library in the pSPORT1 vector (Invitrogen) 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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