title: GSM7135602: VUILD67-LF; Homo sapiens; RNA-Seq
library_construction_protocol: Tissue samples were digested in either collagenase I/dispase II (1 μg/ml) or Miltenyi Multi-Tissue Dissociation Kit using a gentleMACS Octo Dissociator (Miltenyi Inc.). Tissue lysates were serially filtered through sterile gauze, 100- and 40-μm sterile filters (Fischer). The resulting suspensions then underwent cell sorting using serial columns (Miltenyi Microbeads, CD235a and CD45) or fluorescence-activated cell-sorting (FACS) at VUMC or FACSat the Translational Genomics Research Institute (TGen). CD45− and C45+ populations were mixed 2:1 in samples processed at VUMC and used to generate the scRNA-seq libraries. At TGen, calcein acetoxymethyl was used to stain live cells, and 10,000 to 15,000 live cells were sorted directly into the 10x reaction buffer and transferred to the 10x 5′ chip A (10x Genomics). scRNA-seq libraries were generated using the 10x Chromium platform 3′ v2 or 5′ library preparation kits (10x Genomics) according to the manufacturer's recommendations and targeting 5000 to 10,000 cells per sample.