title: GSM8604624: par-cHSF1_37_dox_2; Homo sapiens; RNA-Seq
library_name: GSM8604624
library_construction_protocol: Cellular RNA was harvested using the RNeasy Plus Mini Kit with QIAshredder homogenization columns (Qiagen). RNA samples were quantified using an Advanced Analytical Fragment Analyzer. High throughput three prime digital gene expression (HT-3'DGE) was adapted from the Soumillon et al. protocol (Soumillon et al. BioRxiv https://www.biorxiv.org/content/10.1101/003236v1) to perform reverse-transcription reactions using a dedicated RT primer containing sample and unique molecular identifier (UMI) sequences, to generate four subpools of 24 samples each. After second strand synthesis, Illumina adapters were added using the Nextera Flex tagmentation protocol substituting the normal P500 primer with an ad-hoc primer. Final libraries were cleaned using SPRI beads at 0.7X and quantified using the Fragment Analyzer and qPCR before being loaded for paired-end sequencing using the Illumina NextSeq500 in high-output paired-end mode (20/57 nt reads).