title: GSM9145564: pool B; Mus musculus; OTHER
library_name: GSM9145564
library_construction_protocol: For CITE-Seq analysis, single cell libraries were generated using the BD Rhapsody system. Single cell suspensions from 3 tumors per group closest to the median were stained for multiplexing and with AbSeq oligo antibodies according to manufacturer's protocol (BD Doc ID: 214419 Rev 2.0) with slight modifications. In detail, multiplexing of samples was achieved by labelling with the Custom BD Single-Cell Multiplexing MHC I Set (BD 626545), followed by equal pooling of 3 samples each (resulting in 2 pooled samples), Fc blocking with TruStain FcX PLUS antibody (Biolegend 156604) and staining with AbSeq oligo antibody mix (see Table S8). Single cell capture and cDNA synthesis was performed with the BD Rhapsody Single-Cell Analysis System and the Cartridge Reagent Kit (BD 633731), the Cartridge Kit (BD 33733) and the cDNA Kit (BD 633773) according to manufacturer's protocol (BD Doc ID: 210966 Rev. 1.0). Libraries were generated with the WTA Amplification Kit (BD 633801) according to manufacturer's protocol “mRNA Whole Transcriptome Analysis (WTA), AbSeq, and Sample Tag Library Preparation” (Doc ID: 23-21752-00). Pooling of libraries of each cartridge was calculated following instructions given in BDs' “SEQ Calculator v1.2.xlsm” and libraries were sequenced (paired-end) on the Illumina platform.   Libraries were generated with the Targeted Amplification Kit (BD, 633774) and the Mouse Immune Response Panel (BD, 633753) according to manufacturer's protocol (Doc ID: 214508 Rev. 3.0). Pooling of libraries of each cartridge was calculated following instructions given in BDs' “SEQ Calculator v1.2.xlsm” and libraries were sequenced (paired-end) on the Illumina platform.
identifiers.namespace: GEO
identifiers.id: GSM9145564_r1