GSM5516188: Dll1- control tumor; Mus musculus; RNA-Seq
Single cell suspensions were generated by digestion with collagenase type IA and hyaluronidase  for 55 minutes at 37oC. Post-whole tumor tissue digestion, tumor cells were brought to single cells using 0.25% trypsin EDTA and Dispase/DNAse solution sequentially. Red blood cells were removed using 0.64% sterile ammonium chloride solution for 5 min at 37 °C. Tumor cells were further passed through a 0.45-μm strainer to obtain single cells. Single cells were stained with a combination of antibodies (biotynated CD31, CD45 and Ter 119), followed by streaptavidin PEcy7. Cells were sorted on Aria II at the Upenn Flow Cytometry Core Laboratory. Dataset consists of a total of two single-cell datasets generated using the 10x Genomics Chromium platform. Prior to GEM generation, the sample's concentration (cells/ul) and viability (%) were determined by staining the cells with trypan blue and by manually counting using an inverted microscope. Next-generation sequencing libraries were prepared following the 10X Single Cell v3 protocol without modification. Sample quality was evaluated using Agilent's Bioanalyzer High Sensitivity reagent kit following cDNA amplification and library construction. The completed libraries were quantified using the Kapa Library Quantification Kit and the Life Technologies Viia7 Real-time PCR instrument. Libraries were diluted to 2nM and sequenced in a paired-end, single-indexed format on the Illumina NovaSeq SP using the Rapid Run v3 chemistry.