Leukemic peripheral blood (PB) samples from five AML patients were collected after obtaining informed consent.Mononuclear cells (MNC) were purified from the samples by Ficoll– Hypaque density gradient centrifugation, resulting in approximately 1–3 108 cells per sample. To determine the CD34 and CD38 content, cells were labeled with anti-human CD34- fluorescein isothiocyanate (FITC) anti-CD38-phycoerythrin (PE) and IgG isotype control monoclonal antibodies (Miltenyi Biotec, Auburn, CA, USA) and the CD34 and CD38 expression levels were determined for each sample using FACS-Calibur (Becton Dickinson, Franklin Lakes, NJ, USA).
Extracted molecule
total RNA
Extraction protocol
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
Label
biotin
Label protocol
Biotinylated cDNA were prepared according to the standard Affymetrix protocol from 600 ng total RNA (Affymetrix wt_sensetarget_label_manual)
Hybridization protocol
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
