GSM8763543: SAM24417675 OF9_day7; Mus musculus; RNA-Seq
RNA was extracted using RNeasy Kits (Qiagen, 74104), and RNA quantity and quality were measured with a Nanodrop (Thermo Scientific). For each sample, 100 ng RNA was used to synthesize cDNA using the iScript™ cDNA Synthesis Kit (BioRad, 1708890). Total RNA was quantified using the Qubit RNA HS Assay Kit (Thermo Fisher Scientific, Q32852), and the quality was assessed with RNA ScreenTape on the 4200 TapeStation (Agilent Technologies, 5067-5576). cDNA libraries were generated from 2 ng RNA using the Smart-Seq V4 Ultra Low Input RNA Kit (Takara, 634894). Subsequently, 150 pg cDNA was employed to create sequencing libraries using the Nextera XT DNA Sample Preparation Kit (Illumina, FC-131-1024). Libraries were quantified using the Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific, Q32851), and average library size was determined with D1000 ScreenTape on the 4200 TapeStation (Agilent Technologies, 5067-5582). Libraries were pooled and sequenced on the NovaSeq 6000 (Illumina, San Diego, CA) to produce an average of 30 million single-end 50-base pair reads per sample.