GSM6876424: 4_Ctrl_PriMono; Homo sapiens; RNA-Seq
The total RNA was extracted from cell lysate in QIAzol® (Qiagen). Briefly, 180 ul chloroform was added to 800 – 900 ul of cell lysate and mixed. The mixture was centrifuged at 12,000 g for 15 minutes at 4 0C. The clear upper aqueous layer was transferred to a new tube and mixed with an equal volume of 70% ethanol. The extraction was continued using the RNeasy MinElu column (Qiagen). The extracted RNA was examined on High Sensitivity RNA Screen Tape (Agilent) before use. The RNA-seq libraries were prepared using SMART-Seq® v4 Ultra® Low Input RNA Kit (Takara Bio) followed by Nextera XT DNA Library Preparation Kit (Illumina) according to manufactures' instructions. Libraries were indexed and sequenced over 1 lane using S4 flowcell of NovaSeq 6000 (Illumina) with 2x100-bp paired end reads.