GSM4743419: skin_E7g; Mus musculus; RNA-Seq
skin grafts and axillary draining lymph nodes (DLNs) were collected from each recipient. Samples from 7 recipient mice were pooled for each genotype and tissue and were digested enzymatically and mechanically. Live CD45+ cells of skin grafts and Live CD45+ TCRβ+ cells of DLNs were sorted from single cell suspensions via flow cytometric activated cell sorting. The 10x Genomics Chromium instrument was used to partition each single cell with barcoded beads. This was followed by generation of single cell barcoded 5' cDNA library using the chromium Single Cell 5' Library & Gel Bead Kit (PN-1000014; 10X Genomics) per the manufacturer's instructions. cDNAs were used for both RNA-seq library generation and TCR V(D)J targeted enrichment followed by V(D)J library generation using the Chromium Single Cell V(D)J Enrichment Kit, Mouse T Cells (PN-1000071; 10X Genomics) per the manufacturer's instructions. Libraries for RNA-seq and V(D)J libraries were sequenced on Illumina NextSeq500 platform using a 150 cycle and 300 cycle High-Output Kit (Illumina), respectively.