title: GSM9191726: Ci10067_Plate_7_C_E01_CD3_low; Homo sapiens; RNA-Seq
library_construction_protocol: Cells were then stained with Aqua LIVE/DEAD stain (Thermo Fisher Scientific), anti-CD3-PacBlue, -CD4-AlexaFluor780, -CD8-AlexaFluor700, -CD14/16/19/56-PerCP-Cy5.5 (dump channel, all separate antibodies), -CD69-APC, -CD137-PE, -CD94-PE-Cy7, and -CD107a-FITC. As per gating strategy shown in Figure S1A, single cell lymphocytes were gated on, followed by live cells (Aqua -), CD4/dump -/-, CD8+ CD3-dim, CD69/137+.. Cells which fit this gating strategy and met the criteria of having a CD3-dim activated response to the given HLA pulsed with KF11 significantly (and 3X) higher than 41A3.CD4 also pulsed with KF11 were then sorted into a 96-well plate containing the lysis buffer previously described using the Aria II FACS sorter. scTCR and scRNA-seq was performed as previously described. Library construction was performed at the Institute for Immunology and Infectious Diseases in Perth, Australia. As in the previous publication, uniquely tagged primers were used for reverse transcription and template switching with a preamplification step to increase yield and transcript length of the single-cell cDNA library. Libraries were then run on separate lanes of an Illumina MiSeq instrument utilizing a 2 x 300 bp paired-end chemistry kit (Illumina Inc.).
identifiers.namespace: GEO
identifiers.id: GSM9191726_r1