Name: Melton normalized human islet 4 N4-HIS1 Library ID: 1898 Organism: Homo sapiens Gender: both Age: 0 Stage: adult Organ: pancreas Tissue: Islets of Langerhans, pooled Host: DH10B Vector: pSPORT1 Vector type: phagemid Insert digest: 5' SalI/NotI 3' Stop Codon Status: without Description: Starting library constructed using SuperScript Plasmid Library kit (Life Technologies) and 5' linker sequences as follows. 5'-TCGACCCACGCGTCCG-3' and 3'-GGGTGCGCAGGC-5'. cDNA made by oligo-dT priming using 5'-PGACTAGTTCTAGATCGCGAGCGGCCGCCCT(15)-3'. Size-selected by column fractionation; average insert size 1.08 kb. Library was amplified once on solid support and plasmid DNA from library was prepared. The library DNA was normalized by method #4 from Bonaldo, Lennon, and Soares (1996 Genome Research 6:791-806); 0.5 microgram single-stranded library plasmid DNA was mixed with 5 micrograms PCR product representing library inserts and hybridized to an Ecot of 20. Single-stranded (unhybridized) plasmids were isolated by hydroxyapatite chromatography and used to make this library. Library arrayed and constructed in the laboratory of D. Melton, Ph.D. (Harvard University).