OE31_HMS_D10_Donor3_Rep6
Cell pellets were resuspended in 350 L Magna Pure External Lysis Buffer (Roche, 06374913001) and stored at -80C. RNA was extracted using the Magna Pure 96 instrument (Roche)  and the Magna Pure 96 Cellular RNA Large Volume kit (Roche, 05467535001). RNA was quantified using the Qubit RNA HS assay (Invitrogen, Cat#Q32852). The RNA integrity Number (RIN) scores were determined by using the TapeStation 2200. For this, the High Sensitivity RNA screen tape (Agilent, Cat# 5067- 5579) and the High Sensitivity RNA ScreenTape Sample Buffer (Agilent, Cat# 5067- 5580) were used, following the manufacturers instructions. RNA library preparation was performed using the Illumina stranded mRNA Prep (Illumina, Cat# 20040534), according to the manufacturer's protocol. A total of 200 ng RNA was used for all libraries as input material. Final Library PCR amplification was done using 12 cycles. Prepared libraries were quantified using the Qubit 1x dsDNA HS kit (Invitrogen, Car# Q33231) and checked for fragment size distribution using the TapeStation 2200 using the D1000 DNA screen tape (Agilent, Cat# 5067-5582) with D1000 Reagents (5067- 5583). The quantified libraries were pooled in equimolar amounts and sequenced on the NovaSeq 6000 system with 2 x 50 bp paired-end (PE) reads, targeting 50 million PE reads per library.