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1st strand cDNA was primed with a Not I - oligo(dT) primer 5'-GACTAGTTCTAGATCGCGAGCGGCCGCCC(T)25-3', double-stranded cDNA was cloned into the NotI and EcoRV sites of pExpress-1 (EcoRV site destroyed upon ligation). Library was size-selected for an average insert size of 1.5 kb. Primary library, non-amplified. Use M13(-21) to generate 3' end reads and M13R to generate 5' end reads. Library was constructed by Open Biosystems (Huntsville, AL). Note: this is a (http://zgc.nci.nih.gov/) Zebrafish Gene Collection library.
Nucleotide
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