title: GSM4189812: Project05_single cells; Mus musculus; RNA-Seq
library_construction_protocol: Primary murine NRASG12V/Mll-AF9 AMLs were harvested from spleens of leukemic mice, stained with antibodies for cell surface markers, and sorted on a FACAria (Becton Dickinson, Franklin Lakes, NJ). Cryopreserved bone marrow aspirates from healthy volunteers were purchased from Lonza (Basel, Switzerland). AML samples with >50% bone marrow blasts were obtained from the University of Minnesota Hematological Malignancies Tissue Bank in accordance with the University of Minnesota Institutional Review Board (IRB #1506E72802 and #0611M96846).Fresh AML bone marrow aspirates were CD34-selected using EasySep Magnet and EasySep Human CD34 antibody kit (StemCell Technologies, Vancouver, BC, Canada). Human AML and normal bone marrow were sorted to isolate CD34+CD38- cells. Sorted or magnet-selected cells were stained with a Live/Dead Viability and Cytotoxicity dye (Life Technologies, Carlsbad, California, # L-3224) and loaded at 250 cells/l for single-cell capture on the C1 Single-Cell Auto Prep Integrated Fluidic Circuit for small cells (5-10 m, Fluidigm, South San Francisco, California). Captured cells were imaged to identify single, live cells for cDNA preparation. SMARTer Ultra Low RNA Kit for the Fluidigm C1 System (Takara Bio, Kusatsu, Shiga Prefecture, Japan) was used to prepare cDNA for sequencing. Fluidigm Reverse Transcription Master mix was used to prepare cDNA for PCR.
attributes.tag: GEO Accession
attributes.value: GSM4189812