BioSample

The UI-R-C0 library is a subtracted library derived from the UI-R-A1 and UI-R-E1 libraries. The UI-R-A1 library consisted of a mixture of individually tagged normalized libraries constructed from rat placenta, adult lung, brain, liver, kidney, heart, spleen, ovary, and muscle. The UI-R-E1 library consisted of a mixture of individually tagged normalized libraries constructed from 8, 12 and 18-day embryo. 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-C0) was constructed as follows: PCR amplified cDNA inserts from a pool of UI-R-A1 and UI-R-E1 clones from which 3' ESTs had been derived was used as a driver in a hybridization with the pooled UI-R-A1 and UI-R-E1 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-C0 library. This procedure has been previously described (Bonaldo, Lennon and Soares, Genome Research 6: 791-806, 1996)