title: GSM8472800: Pooled cells with thymus specific, islet specific, and HEL specific T cells_replicate 2; Mus musculus; RNA-Seq
library_construction_protocol: Tissues were mechanically meshed through 70um strainer to generate single cells suspensions Libraries for single-cell gene expression and sequencing were prepared at the McDonnell Genome Institute (MGI) at Washington University using the 3v3.1 methods. Specifically, cDNA was prepared after the GEM generation and barcoding, followed by the GEM-RT reaction and bead cleanup steps. Purified cDNA was amplified for 11-13 cycles before being cleaned up using SPRIselect beads. Samples were then run on a Bioanalyzer to determine the cDNA concentration. GEX libraries were prepared as recommended by the 10x Genomics Chromium Single Cell 3' Reagent Kits User Guide (v3.1 Chemistry Dual Index) with appropriate modifications to the PCR cycles based on the calculated cDNA concentration.