bulkRNA-seq of HeLA cell : Fucci
Protocols: HeLa/Fucci(SA)2 cells (RCB2867, Riken BRC) were maintained in Dulbeccos Modified Eagle Medium (Nacalai Tesque) containing 10% fetal bovine serum (gibco) and 1% penicillin-streptomycin (Sigma-Aldrich). Bulk RNA was extracted from confluent HeLa/Fucci cells in a 100 mm dish (Thermo ScientificTM, 130182) using the PARIS kitTM (InvitrogenTM, AM1921). The NanoDropTM Lite spectrophotometer (Thermo ScientificTM, ND-LITE) was utilized to quantify the RNA concentration. A calibration curve was generated from the dilution series of extracted RNA to estimate the quantity of extracted RNA based on the obtained Ct values of GAPDH. For gene expression analysis, 1-Step RT-qPCR using TaqPathTM was conducted in an experimental system with a total volume of 16 L. Additionally, the diluted RNA samples were utilized for bulk RNA-seq.