title: RNA-seq analysis of xenograft model to characterize the CNS leukemia
design: Leukemic cells (0.2-1.0x106) were injected into non-pretreated NOD-SCID IL2Rgnull (NSG) mice through the tail vein. Leukemic cells in organs are harvested as follows. After as much peripheral blood was removed with PBS perfusion (Perista Pump, Atto), the organs were removed and mechanically dispersed. Mononuclear cells were isolated by Ficoll-Hypaque density gradient centrifugation. To purify the leukemic cells, mononuclear cells were separated using human CD19 microbeads (Miltenyi Biotec). RNA was extracted from leukemic cells using TRIZOL regent (Thermo Fisher SCIENTIFIC) and cDNA libraries were generated with TruSeq® RNA Sample Preparation Kits v2 (illumina) and sequenced on  Illumina® NextSeq 500 (illumina).