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Five libraries representing EI0.5/12.5 pancreatic bud, E16.5 pancreas, newborn pancreas, adult pancreas, and adult islets of Langerhans were seperately constructed using SuperScript Plasmid Library kit (Life Technologies). cDNA was made by oligo-dT priming and size-selected by column fractionation. 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.
Nucleotide
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