GSM9228174: 11-4_S387 Lung tumour. Vehicle (peanut oil); Mus musculus; RNA-Seq
Lungs from SPC-Cre+ KM genetically engineered mice were microdissected under a stereomicroscope. For obtaining a single cell suspension of tumour total cells, enzymatic dissociation with 1 mg/ml Collagenase D (Roche) and 25 mg/mL DNAse I (Thermo Fisher Scientific) was carried out using the gentleMACS Octo Dissociator (Miltenyi Biotec). Then, tumour suspensions were mashed through a 70 m filter and live cells were enriched using debris removal solution (Miltenyi Biotec). For single-cell RNA-Seq, 40,000 cells were loaded per channel onto a Chromium Chip G using reagents from the 10x Genomics Chromium Single Cell 3′ v3 chemistry according to the manufacturer's instructions. Libraries were constructed following the standard 10x Genomics Chromium 3 prime protocol. Library quality and fragment size distributions were evaluated using the Agilent Bioanalyzer High Sensitivity DNA kit. The prepared scRNA-seq libraries were then sequenced on an Illumina NovaSeq 6000 platform using paired-end 150 bp reads.