title: NextSeq 550 paired end sequencing; RNA-Seq analysis of resting and activated mouse Tregs
library_construction_protocol: Regulatory T cells were induced from fresh prepared mouse CD4 T cells from spleen and lymph nodes Total RNA from rested and activated regulatory T cells (four biological replicates) were extracted using Qiagen RNeasy Mini kit according to the manufacturer's instructions and quantified using the Qubit 2.0 Fluorometer (Thermo Fisher Scientific Inc, #Q32866) and the Qubit RNA BR assay kit (#Q10210). RNA sequencing libraries were prepared from 500 ng of each total-RNA sample using the NEBNEXT Ultra II Directional RNA Library Prep kit with Poly-A mRNA magnetic isolation (NEB #E7490) according to the manufacturer's instructions.