GSM8124603: Don3 OM900 8hr; Homo sapiens; RNA-Seq
Total RNA was extracted from sorted cells with a RNeasy plus mini kit (Qiagen) according to the manufacturer's instructions. The quantity and quality of the isolated RNA was determined with a Qubit (1.0) Fluorometer (Life Technologies) and a 4200 TapeStation System (Agilent). NEBNext Ultra DirectionalRNA Library Prep Kit for Illumina (New EnglandBioLabsInc.) was employed with theNEBNextMultiplex Oligos for Illumina (New EnglandBioLabsInc.) andNEBNext®Poly(A) mRNA Magnetic Isolation Module for cDNA synthesis with the addition of barcode sequences. The sequencing of the pre-pools was performed using the NextSeq2000 (Illumina, San Diego, CA, USA) with the P2 reagents kit V3 (100cycles; Illumina).