GSM8722236: KPC_CCR1 KO2 PDA; Mus musculus; RNA-Seq
To prepare tissues for sequencing, samples were disrupted to single cell suspension by mechanically mincing the tissue, followed by enzymatic digestion with Collagenase V (1 mg/mL in RPMI, #C9263, Sigma-Aldrich) at 37°C for 30 min under constant shaking. After digestion, suspensions underwent serial filtration through 500 µm, 100 µm, and 40 µm mesh cell strainers. We then enriched for live cells using the MACS Dead Cell Removal Kit (#130-090-101, Miltenyi Biotec). The resulting single cell suspensions were brought to the University of Michigan Advanced Genomics Core for cDNA library preparation and sequencing. Samples were run using 50-cycle paired-end reads on the NovaSeq 6000 (Illumina) to a depth of 100,000 reads.