GSM7597867: PD CD8 TEMRA; Homo sapiens; RNA-Seq
Samples of different participants of the same group (four HC or five PD) for each sorted subset were pooled and then centrifuged at 4°C 400g for 10 minutes. Discard supernatant, keep cells in 100ul 0.5% BSA/PBS (without Ca2+/Mg2+) at 4°C and proceed to single cell isolation and sequencing. Cell quantification and viability wasassessedusing the C-Chip Disposable Hemocytometer from NanoEnTek. Cells with a viability above 90% werethenresuspended in PBS at afinalconcentration of 1000 cells per μL. The entire procedure was carried out on ice whenever possible. Single cells were processed using the Chromium Next GEM Single Cell 3ʹ and Library Kits v3.1 from 10xGenomics, following the manufacturer's protocol. Libraries were purified using SPRIselect magnetic beads from Beckman Coulter, and their quality was evaluated using High Sensitivity D5000 ScreenTape Assay on an Agilent 4200 TapeStation device. Single-cell libraries with fragment sizes ranging between450-580 bp were sequenced on Illumina NovaSeq 6000system using theNovaSeq 6000 S1 Reagent Kit v1.5 (200 cycles).