GSM9611324: hTERT-RPE1 cells, knockdown ccnd1, dox, 360, rep2; Homo sapiens; OTHER
At each collection timepoint, the cells were washed once with PBS, and lysed with TRIReagent. RNA quality control, library preparation, sequencing and initial data processing were performed by Lexogen NGS Services, Lexogen GmBH, Austria. In brief, samples were characterized by UV-Vis spectrophotometry (Nanodrop 2000c, ThermoFisher) and RNA integrity was assessed on a Fragment Analyzer System using the DNF-471 RNA Kit (15nt) (Agilent). The libraries were constructed using Lexogen's QuantSeq 3' mRNA-Seq Library Prep Kit FWD V2. RNA input to the library preparation was normalized to 75ng across all samples. Prepared libraries were quality controlled on a Fragment Analyzer System using the DNF-474 using the HS-DNA kit (1-6000bp) (Agilent). cDNA libraries were sequenced with an Illumina NovaSeq X platform using 100nt single end read length, with 40 million reads on average per sample.