GSM6381778: MC38_wt_BM_blood, mRNA-derived cDNA; Mus musculus; RNA-Seq
Single-cell suspensions were stained with CD45 antibody and viability dye, then live CD45+ cells sorted by flow cytometry. Individual sorted cell samples were stained using the TotalSeq-B hashtag antibodies (Biolegend) according to the manufacturer's specifications and the combined cells were stained using Biolegend's TotalSeq-B protein-targeted antibodies according to the manufacturer's protocol. Stained cells were subjected to library preparation using the 3' single-cell dual index transcriptome kit (10X Genomics) targeting 30,000 total cells. For bulk RNA-seq experiments, cells sorted by flow cytometry were subjected to RNA isolation using the Arcturus PicoPure kit (Thermo Scientific) followed by library preparation targeting polyA+ RNA. The Chromium Single Cell 3' Reagent Kit v3.1 (Dual Index) (10X Genomics) was used according to the manufacturer's specifications. Bulk RNA-seq libraries were constructed using the SMARTer v4 kit (Takara Bio). mRNA-derived libraries were constructed independently; a combined library containing hashtag and antibody dependent tag oligos was also constructed (ADT_library). For bulk RNA-seq experiments, polyA+ RNA was converted to cDNA, subjected to amplification and library preparation using SMARTer v4 (Takara) for Illumina single-end sequencing. CITE-seq (Cellular Indexing of Transcriptomes and Epitopes by sequencing)