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Starting library constructed using SuperScript Plasmid Library kit (Life Technologies). cDNA made by oligo-dT priming. 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.
Nucleotide
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