GSM6862357: Mouse_scRNA-seq_TDLN_APC; Mus musculus; RNA-Seq
Cell sorting was performed byFACS Aria II (BD Biosciences). For scRNA-seq (10x) of mouse samples live, CD3-CD19-Ly6G+ I-A/I-E+ cells were sorted from naïve LN, tumor-draining LN, and TRAMPC1-GP tumors. For human scRNA-seq (10x) fresh kidney tumors were used, antigen-presenting cells were sorted as live, CD45+, CD3-, HLADR+ Single cell RNAseq libraries were generated using the Chromium Single Cell 5'1007Library & Gel Bead Kit (10x Genomics) according to the manufacturer's protocol. Briefly, cells were sorted and captured into the Gel Beads-in-emulsion (GEMs). After reverse transcription, the GEMs were disrupted and the barcoded cDNA was isolated, pooled, and amplified by PCR (13cycles). The amplified cDNA was fragmented, and subjected to end repair and A-tailing followed by a sample index PCR (16 cycles). The purified libraries were sequenced to a depth of 50,000 reads/cell on a 1013HiSeq3000 (Illumina) with 26 cycles for read 1, 8 cycles for index 1 (i7), and 91 cycles for read 2.