GSM9290768: 30 °C CD4+ T cells, GFP-, replicate 1; Mus musculus; RNA-Seq
RNA from cells was isolated using the mirVANA RNA isolation kit. To measure the library concentration, NEBNext Ultra II Directional RNA Library Prep kit  (New England BioLabs) was used. To enrich polyA RNA for library preparation, NEBNext Poly(A) mRNA Magnetic Isolation Module (New England BioLabs, Ipswich, MA) was used with 1 µg  total RNA as input. Next, NEBNext Ultra II Directional RNA Library Prep kit (New England BioLabs) was used for unique dual index library preparation under PCR cycle number of 8. After library QC and Qubit quantification (ThermoFisher, Waltham, MA), the normalized libraries were sequenced using NextSeq 2000 Sequencer (Illumina, San Diego, CA). Under the setting of PE 2x61 bp, ~45M reads were generated for each sample.