GSM9652395: PAH PASMC4 Ctrl-siRNA; Homo sapiens; RNA-Seq
Total RNA was isolated using RNeasy Kit (Qiagen) catalog#74104 following the manufacturer's protocol. Libraries for RNAseq analysis were constructed using the Lexogen 3' kit according to manufacturers'protocols (Lexogen, Austria), and validated using Qubit 4 (Thermofisher Scientific). Successful libraries were pooled and diluted to an equimolar concentration resulting in a total pool of 4 nM. Dilution and denaturationwere performed according to Illumina protocols to result in a 2.2pM input (optimized for the Lexogen 3' kit). Single-end sequencing of 75 cycles was performed on an Illumina Nextseq550 using a High Output v2 reagent cartridge with each sample having a minimum depth of 5 million reads.