BioSample

The UI-R-E1 library is a subtracted library derived from the UI-R-E0 library. The UI-R-E0 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-E1) was constructed as follows: PCR amplified cDNA inserts from a pool of UI-R-E0 clones from which 3' ESTs had been derived was used as a driver in a hybridization with the UI-R-E0 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-E1 library. This procedure has been previously described (Bonaldo, Lennon and Soares, Genome Research 6: 791-806, 1996)