title: GSM8395067: Patient_4 pre-AZA; Homo sapiens; RNA-Seq
library_name: GSM8395067
library_construction_protocol: Bone marrow mononuclear cells (BMMCs) were isolated from BM samples using Ficoll-Paque Plus (GE Healthcare, Massachusetts, US) density gradients per manufacturer's instructions. CD34+ cells were isolated by immunomagnetic separation using an autoMACs Pro Separator (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) following the manufacturer´s instructions. Sorted cells were snap-frozen and stored at -80ºC. Total RNA was extracted using the RNeasy Micro Kit (Qiagen, Valencia, CA, USA) following the manufacturer's protocol. RNA integrity was assessed on an Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) using the RNA 6000 Nano Assay kit. RNA quantities were assessed by qubit 2.0 (ThermoFisher Scientific, Massachusetts, U.S.A.) using an RNA kit (high/broad sensitivity). Libraries were prepared using the SMART-Seq v4 Ultra Low Input RNA kit (Clontech, California, U.S.), and sequenced on a HiSeq400 sequencer (Illumina) according to manufacturer´s protocol with a read length of 2 × 150 nucleotides.