title: GSM9051257: 21E24_5HT_48h_CTB; Homo sapiens; RNA-Seq
library_construction_protocol: Cells were lysed in QIAzol, and RNA was isolated using the miRNeasy Mini Kit (Qiagen), also following the manufacturer's protocol. An on-column DNase digestion was performed. RNA Seq Library Prep: Using the Kapa RNA HyperPrep Kit with RiboErase (KR1351, Roche Diagnostics, Indianapolis, IN), rRNA was depleted starting from a normalized input of total RNA by hybridization of rRNA to complementary DNA oligonucleotides, followed by treatment with RNase H and DNase to remove rRNA duplexed to DNA. Samples were then fragmented using heat and magnesium. 1st strand synthesis was performed using random priming. 2nd strand synthesis incorporated dUTPs into the 2nd strand cDNA. Adapters were then ligated, and the library was amplified. Strands marked with dUTPs were not amplified allowing for strand-specific sequencing. Indexed libraries that meet appropriate cut-offs for both quantity and quality were quantified by qRT-PCR using a commercially available kit (KAPA Library Quant Kit, Roche Diagnostics, 07960298001) and insert size distribution determined with an Agilent Bioanalyzer (Agilent TapeStation 4200). Samples with a yield of ≥0.5 ng/µl were used for sequencing.