title: GSM6669734: Tcrm_Control_5dpi_VDJ; Mus musculus; RNA-Seq
library_construction_protocol: Heart and medistinal lymph nodes were collected from control adult mice at 5 days post myocardial infarction surgery. All tissues were kept in HBSS/BSA 0.5% at 4°C until processing. Heart samples were digested in collagenase II 1000U/mL for 30 minutes, at 37ºC. After digestion, both tissues were grinded through a 30um mesh and seeded in a plate for FACS staining.Before hashtag staining, samples were labeled with Myhca614-629 tetramer construct as described in the methods section. Afterwards, each condition (e.g Heart, 5d, MI) was stained with a different Hashtag TotalSeq C antibody to enable multiplexing plus the TotalseqC anti-APC antibody to detect tetramer positive cells. T-helper cells were defined as live, lineage (CD11b,CD8a,CB220) negative and CD4+  events and were FACS sorted cells pooled heart and MedLN samples. After sorting, cells were combined at different ratios for single cell RNA sequencing application. Cells were loaded in the 10X Genomics Chromium with the aim of recovering 30,000 cells. Libraries were generated with the Chromium Next GEM Single Cell 5' Reagents Kit v2 Chemistry according to manufacturer ´s instructions.