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cDNA library: Melton normalized mixed mouse pancreas 1 N1-MMS1

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Information for 1 library

pancreas

   1. Melton normalized mixed mouse pancreas 1 N1-MMS1

Melton normalized mixed mouse pancreas 1 N1-MMS1

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).

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