GSM9416732: Fibroblasts, DMSO, repeat 2; Homo sapiens; RNA-Seq
Total RNA extraction and purification was performed using the RNeasy Mini Kit (QIAGEN). Removal of residual gDNA was performed using the TURBO DNA-free Kit (Invitrogen, Waltham, MA, USA). Total RNA (500 ng) was used for library preparation. Enrichment of polyadenylated RNA was performed with KAPA mRNA Capture Kit (Roche, Switzerland) and libraries were prepared using the KAPA RNA Hyper Kit (Roche, Switzerland), according to the manufacturer protocol. The library underwent a final cleanup using the KAPA HyperPure Beads (Roche, Switzerland) after which the libraries' size distribution and quality were assessed using a high sensitivity DNA chip (Agilent Technologies, USA). Libraries were pooled equimolarly and diluted to a final concentration of 750 pM. Sequencing was carried out on a NextSeq 1000 platform (Illumina, USA) using the NextSeq 1000/2000 P2 Reagents kit (200 Cycles) v3, with 4% PhiX (Illumina, USA) added as an internal control.