GSM9542272: LN229,RM6,1.56nM,rep1; Homo sapiens; RNA-Seq
RNA was extracted from cells using Quick-RNA microprep kit (Zymo # R1050) according to the manufacturer's instructions 3' RNA-seq libraries were generated using a QuantSeq FWD Kit (Lexogen # 015) with UMI add-on module according to the manufacturer's instructions. Briefly, 350 ng of input RNA was used for first-strand cDNA synthesis with an oligo(dT) primer, RNA was removed, second strand synthesis was performed with random primers containing unique molecular identifiers, and the generated cDNA purified using magnetic beads. Purified cDNA was amplified by 16 cycles of PCR to generate sufficient material for sequencing and to introduce complete sequencing adapters. Amplified libraries were subject to a further purification step and then assessed for yield and composition on an Agilent Bioanalyzer. Samples were pooled equimolarly, and pooled libraries were subsequently sequenced on an Illumina NextSeq550 platform using a High Output Reagent Cartridge v2 (Illumina # 15057934) in single-end 65 bp configuration.