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The UI-R-Y0 library is a subtracted library derived from an individually-tagged normalized whole-eye (minus the lens) library. The driver for the subtraction consisted of a pool of all previous libraries (UI-R-A0, UI-R-A1, UI-R-E0, UI-R-E1, UI-R-C0, and UI-R-C1). The tag is a string of 3-5 nucleotides present between the Not I site and the oligo-dT track which allows identification of the library of origin of a clone within the mixture. The subtracted library (UI-R-Y0) was constructed as follows: PCR amplified cDNA inserts from previous library clones from which 3' ESTs had been derived were used as a driver in a hybridization with the normalized whole-eye library in the form of single-stranded circles. The remaining single-stranded circles (subtracted library) was purified by hydroxyapatite column chromatography, converted to double-stranded circles and electroporated into DH10B bacteria (Life Technologies) to generate the UI-R-Y0 library. This procedure has been previously described (Bonaldo, Lennon and Soares, Genome Research 6: 791-806, 1996)
Nucleotide
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