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The UI-R-C3 library is a subtracted library of a series, ultimately derived from a mixture of individually tagged normalized libraries from rat placenta, adult lung, brain, liver, kidney, heart, spleen, ovary, muscle, and 8, 12 and 18-day embryos, after a series of subtractions to reduce the representation of cDNAs from which ESTs had already been generated. The following serially subtracted libraries were generated in this process: UI-R-C3, UI-R-C2p, UI-R-C1, UI-R-C0, UI-R-A1, UI-R-E1. The tag is a string of 3-5 nucleotides present between the Not I site and the oligo-dT track whichallows identification of the library of origin of a clone within themixture. The subtracted library (UI-R-C3) was constructed as follows: PCRamplified cDNA inserts from UI-R-C2p clones from which 3' ESTs had been derived was used as a driver in a hybridization with the UI-R-C2p 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 (LifeTechnologies) to generate the UI-R-C3 library. This procedure has been previously described (Bonaldo, Lennon and Soares, Genome Research 6:791-806, 1996)
Nucleotide
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