title: GSM4732979: mRNA; Mus musculus; RNA-Seq
library_construction_protocol: CD11b+ cDCs from C57BL/6 mice inhaled house dust extract together with ovalbumin were purified by flow cytometry, and stained with Total A-Seq oligo-conjugated antibodies against CD11b and Ly-6C according to manufacturer's instruction (BioLegend). The cells were counted and examined for viability using a TC-20 cell counter (Bio-Rad). 7000 live cells at 7×105 cells/mL concentration with 98% viability were loaded into the Single Cell Chip followed by forming single cell emulsion in Chromium Controller (10x Genomics). The cDNA for antibody derived transcripts (ADT) and gene derived transcripts was generated and amplified according to manufacturer's instructions (10x Genomics and BioLegend). The Gene Expression library and the ADT Library were prepared using the Chromium single cell 3′ v2 library and gel bead kit (10x Genomics) and additional reagents recommended in the protocol of Total A-seq (Biolegend). The two libraries were mixed at 10:1 molar ratio (Gene Expression library to ADT library).