title: Illumina HiSeq 4000 paired end sequencing; RNA-Seq of conventional and regulatory CD4+ T cells of healthy human donors
design: RNA-Seq of conventional and regulatory CD4+ T cells of healthy human donors
library_construction_protocol: 2000 cells were sorted as 7-AAD-CD3+CD4+CD127-CD25high Treg cells and   7-AAD-CD3+CD4+CD127varCD25var effector T cells from human PBMCs  blood collection straight ex vivo flow sorting none applied modified SMART-Seq2: Agencourt RNAClean XP paramagnetic beads (Beckman Coulter) were used in combination with a DynaMag-96 side skirted magnet (Thermo Fisher). cDNA was generated with SuperScript II Reverse Transcriptase Kit (Thermo Fisher) and amplified with HiFi HotStart PCR Mix (KAPA Biosystems). For DNA clean-up Agencourt AMPure XP beads were used (Beckman Coulter) as above.  Nextera XT DNA sample preparation and index kits (Illumina) were used for preparation of libraries
attributes.tag: Experimental Factor: cell type
attributes.value: CD4-positive, alpha-beta T cell