GSM5412807: CTR rep2; Mus musculus; RNA-Seq
Mice were sacrificed, tumours isolated and processed in order to obtain cell suspensions. Two samples/group (each one composed by a pool of 4 mice), were enriched for CD45 marker on FACS ARIA (BD biosciences) instrument. FACS-sorted cells were resuspended in RPMI with 5% FBS to a final concentration of 1000 cells per μl and processed using the 10x Genomics Chromium Controller and the Chromium NextGEM Single-Cell 3′ GEM, Library & Gel Bead kit v3.1 (Pleasanton, California, United States) following the standard manufacturer's instructions. In brief, 10,000 live cells were loaded onto the Chromium controller to recover 4,000 single cell GEMs per inlet uniquely barcoded. After synthesis of cDNA, sequencing libraries were generated. Final 10X library quality was assessed using the Fragment Analyzer High Sensitivity NGS kit (Agilent Technologies, Santa Clara, CA, USA) and then sequenced on the Illu-mina NextSeq500 (Illumina, San Diego CA, USA). After synthesis of cDNA, sequencing libraries were generated. Final 10X library quality was assessed using the Fragment Analyzer High Sensitivity NGS kit (Agilent Technologies, Santa Clara, CA, USA) and then sequenced on the Illu-mina NextSeq500 (Illumina, San Diego CA, USA).