title: RNAseq of human U937 cell line treated with PMA for 96 hrs: replicate 2
design: The cytoplasmatic rRNA removal was performed on total RNA using the Ribo-Zero rRNA removal Kit (Epicentre, Madison, WI, USA). The stranded-oriented RNA-seq library was prepared using the TruSeq Stranded Total RNA Sample Prep Kit (Illumina, San Diego, CA, USA). Briefly, each RNA was chemically fragmented prior to the random priming reverse transcription reaction for first strand cDNA generation. The fragmentation step resulted in a RNA-seq library including inserts ranging in size from approximately 100-400 bp. During the second strand synthesis, dUTP was incorporated in place of dTTP, thus preventing amplification of this strand during the subsequent PCR step and retaining strand information. cDNA libraries were sequenced on the Illumina Next Seq 500 platform. Paired-end reads of 100 nt were generated for each fragment.