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RNA was prepared from a pool of 6 anonymous Wilms' tumor RNAs. RNA was prepared by acid-phenol, followed by one round of oligo dT selection. cDNA library preparation was with the BRL/Life Tech. Superscript Plasmid system. An oligo-dT NotI primer for first strand synthesis generated gcggccgccc(t)n at the 3' end of the clones. A 5' SalI adaptor was used with sequence 5'-gtcgacccacgcgtccg-3'. Resulting cDNAs were size selected (average size 2 kb), NotI digested, and ligated into NotI/SalI-cut pSPORT1. Library was constructed by Dr. Manfred Gessler.
Nucleotide
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