GSM5960562: MO-04-CMP; Homo sapiens; RNA-Seq
total RNA was extracted from mononuclear cells or sorted HSPC (1000 to 10000 cells) using the RNeasy Mini or Micro Kit (Qiagen). Libraries were prepared with TruSeq® Stranded Total RNASample preparation kit (Illumina) according to supplier's recommendations. Briefly the ribosomal RNA fraction was removed from 1μg of total RNA using the Ribo-Zero™ Gold Kit (Epicentre). Fragmentation was then achieved using divalent cations under elevated temperature to obtain approximately 300bp pieces. Double strand cDNA synthesis was performed using reverse transcriptase and random primers, Illumina adapters were ligated and cDNA library was PCR amplified for sequencing. Paired-end 75b sequencing was then carried out on a HiSeq4000 (Illumina).