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The library was made from oligo(dT) primed cDNA and cloned into the pCMV-SPORT6 vector (Invitrogen Corp., Carlsbad, CA). PolyA RNA was primed with an oligo dT primer (5'-GACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'), ligated to a SalI adapter (5'-TCGACCCACGCGTCCG and 5'- CGGACGCGTGGG) and digested with NotI. cDNA was size selected from 0.6-1.2kb using 1.1% agarose gel electrophoresis then ligated into NotI and SalI-digested pCMV-SPORT6 vector. The library was constructed at DOE Joint Genome Institute (Walnut Creek, CA).
Nucleotide
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