Name: Melton normalized mixed mouse pancreas 1 N1-MMS1 Library ID: 1948 Organism: Mus musculus Strain: ICR Gender: both Age: 0 Stage: mixed Organ: pancreas Tissue: pool of whole pancreas, pancreatic bud, Islets of Langerhans Host: DH10B Vector: pSPORT1 Vector type: phagemid Insert digest: 5' SalI/NotI 3' Stop Codon Status: without Description: Five libraries representing EI0.5/12.5 pancreatic bud, E16.5 pancreas, newborn pancreas, adult pancreas, and adult islets of Langerhans were separately constructed using SuperScript Plasmid Library kit (Life Technologies). cDNA was made by oligo-dT priming using primer 5'-PGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3' and size-selected by column fractionation. Cloning was accomplished via 5' linkers: 5'-TCGACCCACGCGTCCG-3' and 3'-GGGTGCGCAGGC-5'. Libraries were amplified once on solid support and plasmid DNA from each library was prepared and mixed in equal amounts. The mixed library DNA was normalized by method #4 from Bonaldo, Lennon, and Soares (1996 Genome Research 6:791-806); 0.5 microgram single-stranded mixed library plasmid DNA was mixed with 5 micrograms PCR product representing mixed library inserts and hybridized to an Ecot of 6. Single-stranded (unhybridized) plasmids were isolated by hydroxyapatite chromatography and used to make this library. Library constructed and arrayed in the laboratory of D. Melton, Ph.D. (Harvard University).