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A complementary DNA (cDNA) library from human erythroid precursor cells was constructed using SMART PCR (polymerase chain reaction) cDNA Library Construction Kit (Clontech, Palo Alto, CA) according to the manufacturer's directions, but with slight modifications. Briefly, reverse transcription was performed in the presence of 1 umol/L peptide nucleic acid (PNA) oligos (N-terminal)-biotin-GTC-CAC-CCG-AAG-CTT-G-(C-terminal) and (N-terminal)-biotin-C(T/C)T-GAA-GTT-CTC-AGG-A-(C-terminal). Synthesized cDNA was digested with SfiI and size-selected on a 1% agarose gel (>800bp). Large-scale sequencing of the library was performed by the NIH Intramural Sequencing Center (NISC; Http://www.nisc.nih.gov/).
Nucleotide
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