title: Mus musculus wild-type B cells activated with LPS,transduced with pMSCV shRNA and GFP, direct RNA sequencing, biological replicate 1
library_construction_protocol: 3 mln cells were collected and the cell pellet was resuspended in TRIReagent (Sigma-Aldrich; cat. no: T9424). RNA was isolated according to the manufacturer's protocol. RNA was then purified using KAPA Pure Beads (Roche; cat. no: KK8001) in 1:1.8 RNA to beads ratio. For direct RNA sequencing, 4.3 µg of total RNA from B cells was mixed with 200 ng of oligo-(dT)25-enriched mRNA from Saccharomyces cerevisiae and standards with predefined poly(A) lengths and used for library preparation with a Direct RNA Sequencing Kit (SQK-RNA002; ONT) as described in Bilska et al. (2020).