GSM6604887: Donor 3, trispecific aNKp46/Fc/aCD20; Homo sapiens; RNA-Seq
Total RNA was extracted using RNeasy micro kit (Qiagen) according to the manufacturer's instructions. RNA concentration was assessed using the Qubit RNA HS Assay kit (Thermofisher). RNA-Seq libraries were prepared from 80ng to 200ng of total RNA using the KAPA RNA HyperPrep kit (Roche). mRNAs were enriched with oligo dT magnetic beads and then fragmented for 5min30 at 94°C. Cleaved mRNA fragments were used as templates for double stranded DNA synthesis followed by A-tailing. The 8 bp UMI containing TruSeq Compatible Duplex Y adapters UDI-UMI, (Unique dual index - Unique molecular identifier) from IDT were then ligated on both sides of the cDNA. After bead-based purification, libraries were amplified by PCR for 10-12 cycles. Cleaned-up libraries were quantified using the Qubit 1X dsDNA HS Assay kit (Thermofisher) and quality was assessed using TapeStation.