GSM4971725: cDNA7044; Homo sapiens; RNA-Seq
Total RNA was isolated from cell lysates using the RNeasy Micro Kit (Qiagen), including on-column deoxyribonuclease digestion, and was analyzed for quality using the RNA 6000 Pico Kit (Agilent Technologies). Poly(A)-enriched next-generation sequencing library construction was performed using the KAPA mRNA Hyper Prep Kit (KAPA Biosystems) with 100  ng of input total RNA and 15 amplification cycles according to the manufacturer's protocol. The quality and quantity of the individual libraries was assessed using the High Sensitivity DNA Kit (Agilent).  The libraries were equimolar pooled and an additional 1X bead clean-up was used to remove adapter dimer. The pooled libraries were quantitated using the KAPA Library Quantification Kit, Universal (KAPA Biosystems) and sequenced on an Illumina NextSeq 500 with paired-end 75-base-pair (bp) read lengths.