RNAseq of Human: breast cancer cells
RNA-seq libraries were prepared using the TruSeq Stranded Total RNA Library Prep Gold (Illumina, 20020599) at the OncoGenomics Shared Resources (OGSR) of Sylvester Comprehensive Cancer Center. In brief, extracted RNAs were quantified and qualitatively assessed on the ThermoFisher QuBit Fluorometers High Sensitivity RNA (Cat #Q32855) and Agilent Fragment Analyzer High Sensitivity RNA kits (Cat #DNF-472-0500), and up to 500 ng of high-quality (RQN > 9.0) RNAs were used as input. Total RNA was first depleted of ribosomal RNA using RiboZero beads, followed by fragmentation, then first and second strand cDNA conversion, and finally library synthesis with 3 adenylation, adapter ligation and PCR amplification. Finished libraries quantified and qualitatively assessed on the ThermoFisher QuBit Fluorometers High Sensitivity DNA (Cat #Q32855) and Agilent Fragment Analyzer High Sensitivity NGS kits (Cat # DNF-474-0500) prior to normalization and pooling in preparation for sequencing.