GSM5101006: +CM HL-60, rep3; Homo sapiens; RNA-Seq
RNA from hematopoietic cells (CD34+) was purified using the RNeasy Mini or Micro kit (both Qiagen, Hilden, Germany) according the manufacturer's instructions. DNase digested total RNA samples used for transcriptome analyses were quantified (Qubit RNA HS Assay, Thermo Fisher Scientific Inc., Waltham, MA, USA) and quality measured by capillary electrophoresis using the Fragment Analyzer and the 'Total RNA Standard Sensitivity Assay' or the Bioanalyzer assay “Eukaryote Total RNA Pico” (Agilent Technologies, Inc., Santa Clara, CA, USA). The mean RNA Quality Number (RQN) of the samples in this study was 8.9. The library preparation was performed according to the manufacturer's protocol using the 'NuGen Universal Plus mRNA-Seq Library Prep Kit' for Illumina®. Briefly, 10 ng total RNA were used for poly(A) RNA selection, fragmentation, cDNA generation, adapter ligation, strand selection and library amplification. Bead purified libraries were normalized and finally sequenced on the HiSeq 3000/4000 system (Illumina Inc. San Diego, CA, USA) with a read setup of 1x150 bp. The bcl2fastq tool was used to convert the bcl files to fastq files as well for adapter trimming and demultiplexing.