title: GSM8633514: TDP43-FL, DMSO, 8hr, replicate 2; Homo sapiens; RNA-Seq
library_name: GSM8633514
library_construction_protocol: Total RNA was harvested from cells using the RNeasy Micro Kit (Qiagen), following the manufacturer's protocol. RNA integrity (RIN) and concentration were measured by High Sensitivity RNA Screen Tape Analysis (Agilent) on a 4200 TapeStation System (Agilent). RNA-Seq libraries were generated using the QuantSeq 3' mRNA-seq Library Prep Kit FWD for Illumina (Lexogen A01173) with high quality total RNA (RINe>8) as input template.  Briefly, total RNA was primed with oligo(dT) for reverse transcription, followed by RNA removal.  Lexogen's UMI Second Strand Synthesis Module was added to introduce unique molecular identifiers to identify and remove PCR duplicates.  Amplified cDNA was purified using magnetic beads followed by 12 cycles of PCR with dual indexes and PCR purification.  Library quantity and quality were assessed with High Sensitivity D1000 ScreenTape (Agilent 5067-5584).  Libraries were pooled in equimolar ratios and the sequencing pool was purified with magnetic beads to remove residual adaptor dimers.  Sequencing reads were generated on an Illumina NovaSeq 6000 instrument (75 bp single end) by SeqMatic (Fremont, CA, USA).