title: GSM2828596: 3030_CD8_ZK_IFNgneg; Homo sapiens; RNA-Seq
library_construction_protocol: Cryopreserved PBMC were thawed, stimulated with ZIKA pool of peptides or anti-CD3/CD28 stimulation or with no stimuli. After 3 hr of stimulation cells were stained with antibodies and IFNg capture assay, 200 cells of CD8 T cells were sorted using BD Aria 3 or BD Aria Fusion cell sorter for each condition, multiple of 200  cells were collected when possible.  CD8 responding and non responding  cells were defined with the expression of the followiing antibodies CD3+CD8+IFNg+ and CD3+CD8+IFNg- respectively. Microscaled cDNA amplification of purified total RNA (1 to 5 ng) following the smart-seq2 protocol (16 cycles of cDNA amplification) (Picelli et al., 2014). cDNA was purified using AMPure XP beads (Beckman Coulter). From this step, 1 ng of cDNA was used to prepare a standard Nextera XT sequencing library (Nextera XT DNA sample preparation kit and index kit, Illumina).
identifiers.namespace: GEO
identifiers.id: GSM2828596