GSM8667742: B16F10 tumors treated with Cisplatin; Mus musculus; RNA-Seq
Following tumor disaggregation, lysates were incubated with TotalSeq-B0301, B0302, B0303 and B0304 anti-mouse hashtag antibodies (Biolegend) and replicates were pooled. At least 50,000 CD45+ cells were sorted from each group into PBS using BD FACSAria II Cell Sorter. Cells were loaded into the Chromium Single Cell Controller with Single Cell 3' Gel Beads and Partitioning Oil to generate Gel Bead-in-EMulsion (GEMs) droplets. Next, mRNA and cell-hashing antibody-derived tags (HTOs) were reverse transcribed, barcoded and amplified according to the manufacturer's recommendations. Amplified cDNA was size separated with SPRI beads (Beckman Coulter) into <300 nt fragments with HTOs and >300nt fragments with mRNA-templated cDNA. cDNA sequencing libraries were generated according to 10X Genomics guidelines. Purified HTOs were amplified to append P5 and P7 adapter sequences compatible with the Illumina Flow Cell using a common SI-PCR forward primer and pooled with the cDNA. Libraries were sequenced on an Illumina NextSeq 500 instrument