GSM9256283: lung metastasis rep2; Mus musculus; RNA-Seq
At humane endpoint, liver and lung metastases were isolated, tissues digested as described above and live RFP⁺ tumor epithelial cells sorted from each metastatic site and subjected to single-cell RNA sequencing, profiling ~10,000 cells per tissue. Sample processing for single-cell RNA-seq was done using Chromium Single Cell 3' Library reagent kit v2 following manufacturer's guide (10x Genomics). Cell density and viability of single-cell suspension were determined by Vi-CELL XR cell counter (Beckman Coulter) or by manual counting using a hemocytometer. We loaded ~10,000 cells on each chip. cDNAs and libraries were prepared following the manufacturer's user guide (10x Genomics). Libraries were profiled by Tapestation D1000 kit (Agilent) and quantified using IQubit HS dsDNA kit (Invitrogen). Each library was sequenced in one lane of HiSeq4000 (Illumina) following the manufacturer's sequencing specification (10x Genomics).