Cancer discoveries could enhance immunotherapy, breastcancer care uva NatureComms aacr
mice were minced into small pieces and incubated for 90 mins at 37 °C in dissociation buffer containing 2 mg/mL of collagenase type I , 1 mg/mL of DNase , and 2.5 units/mL of neural protease . The digested tissues were separated into a single-cell suspension by passing through a 100 μM cell strainer. Single-cell isolates were stained with primary antibodies for 30 mins on ice, followed by secondary antibodies, and then washed using FACS buffer.
After library preparation, the FASTQ files were mapped using the CellRanger pipeline to generate a count matrix. The matrix generated by the CellRanger pipeline was imported into the Seurat R package for performing the additional QC and in-depth analysis. In summary, cells that had low RNA counts were filtered out. The cells were also filtered based on <10% mitochondrial genes.
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