MinION sequencing; Nanopore RNA-seq of chromatin RNA sequencing in human primary Naive CD4+ T cells
naive CD4+ T cells have been isolated from peripheral PBMCs by density gradient centrifugation with Ficoll-Paque Plus. From PBMCs, T cells were negatively selected with magnetic separator (AutoMACS). Naive CD4+ were FACS sorted as CD4+CD25-CD127-/highCD45RO-. Chromatin RNA have been extracted with Maxwell RSC miRNA Tissue kit  Libraries were prepared starting from 75 ng of chromatin RNA, using the PCR-cDNA Barcoding Kit SQK-PCB109 (Oxford Nanopore Technologies, UK) and following manufacturerâ€™s guidelines