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Two directionally cloned cDNA libraries were made from a hematopoietic stem cell-supporting stromal cell line (AFT024) and from a non-supporting stromal cell line (2018). Subtractive hybridization was performed by hybridization of the target, AFT024, single stranded cDNA library in pSport1 to biotinylated RNA transcribed from the driver, 2018 cDNA library in pSport2 with inserts cloned in the complementary orientation. The AFT024-subtracted library contains 4.2x105 clones and is depleted of common housekeeping gene products eg. beta-actin and enriched for transcripts specific to AFT024. For detailed protocols and additional information please see our website at http://stromalcell.princeton.edu.
Nucleotide
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