title: GSM7139421: LN 6 hours post-vaccination; Mus musculus; RNA-Seq
library_construction_protocol: We isolated lymph nodes from mice that were either untreated or 6 hours after subcutaneous/intradermal vaccination with 200 mg of ovalbumin peptide and 20 mg CpG 1826 adjuvant. Lymph nodes were dissociated using Collagenase IV, 0.8 mg/ml Dispase, and Dnase I in RPMI at 37°C and cells using a protocol that is optimized for myeloid and stromal cell recovery (Fletecher et al., 2011). After harvesting, cells were put on ice, washed with PBS/2mM EDTA/0.5% biotin-free bovine serum albumin, and filtered through a 70µm cell strainer. T and B cells were isolated with MACS columns using biotinylated anti-CD3 and anti-CD19 antibodies. A small fraction of the CD3+ or CD19+ cells was pooled with CD3-/CD19- cells for more balanced representation of immune lineages prior to performing scRNA-seq (10X genomics). 10X Genomics 3' v2 was performed according to the manufacturer's instructions