GSM8247669: Respiratory mucosa, IAV, 5dpi, gene expression; Mus musculus; RNA-Seq
Tissue was harvested by microdissection and digested in RPMI (w/ FBS, HEPES, and P/S) for 20 min (LNG) or 30 min (RM/OM). Following enzyme inactivation with EDTA and filtration through a 70 µm membrane, ACK lysis was performed on ice for 1min. Epithelial cells were partially depleted using Biotin Binder Dynabeads pre-loaded with anti-biotin anti-EpCAM antibody during a 20min incubation on ice. Following bead removal, cells were stained with TotalSeqB Hashtag antibodies for 20 minutes before washing, counting, and combining. Finally, cells were captured and processed on either a 10x Chromium or X controller using the 3' transcriptome v3.1 kit and feature barcoding kits. Libraries were constructed as outlined in the 10x Genomics guide for Dual-indexed 3' transcriptomics v3.1 with Feature Barcoding.