title: GSM3011854: Primary TNBC infiltrating T cells case 2; Homo sapiens; RNA-Seq
library_construction_protocol: Primary tumour tissues were resected from two patients with triple-negative breast cancer.  Tissues were treated with collagenase and mechanically dissociated. Single cell suspensions were generated and the viable cells were FACS sorted for CD3+ T cells on a BD FACSAria II Cell Sorter (BD Biosciences). Sorted cells were then counted, assessed for viability and resuspended. Cell suspensions were loaded onto a Chromium Single Cell Chip along with the reverse transcription (RT) mastermix and single cell 3' gel beads, aiming for 2000 – 6000 cells per channel. Following generation of single cell gel bead-in-emulsions (GEMs), reverse transcription was performed using a C1000 Touch Thermal Cycler with a Deep Well Reaction Module (Bio-Rad Laboratories, Hercules, CA, USA). Amplified cDNA was purified using SPRIselect beads (Beckman Coulter, Lane Cove, NSW, Australia) and sheared to approximately 200bp with a Covaris S2 instrument (Covaris, Woburn, MA, USA) using the manufacturer's recommended parameters. Sequencing libraries were generated with unique sample indices (SI) for each sample. Libraries were sequenced on an Illumina HiSeq 2500 High Output Mode using V4 clustering and sequencing chemistry to achieve paired-end 100 bp reads..
identifiers.namespace: GEO
identifiers.id: GSM3011854