title: GSM4761001: Tolerized kidney rep 1; Mus musculus; RNA-Seq
library_construction_protocol: Kidney allografts from untreated and tolerized WT B6  recipients were retrieved on d15 post-transplant, perfused by cold PBS and a single cell suspension was prepared using 'multi tissue dissociation kit 2' (Miltenyi  Biotec, Auburn CA) as per manufacturer's protocol. Naïve B6 kidneys retrieved from WT B6 mice were similarly processed to serve as a control. Cell suspension was subjected to RBC lysis followed by washing. Cell viability was confirmed to be >90%, and cell concentration was adjusted to 2,000 cells/μl. In brief, 10X Genomics microfluidic platform combines single cells with barcoded gel beads containing essential components for cDNA preparation including barcodes to tag individual transcripts from the cells, a Unique Molecular Identifier (UMI) to identify PCR products, and primers for amplification of resulting cDNA. cDNA libraries were checked for quality using Agilent D5000 Tape Station analyzer. cDNA libraries of ~400bp size were subjected to Illumina NextSeq 550 sequencer using NextSeq500/550 High Output Kit v2.5 (150 Cycles).