title: GSM4705884: NALT-Control and Treated cells; Mus musculus; RNA-Seq
library_construction_protocol: We harvested the NALT from the immunized animals at the time of euthanasia and converted them into single-cell suspensions. We subsequently washed the single-cell suspensions with PBS, resuspended them in PBS + 2% FBS, and added 50 nM Helix NP Blue (BioLegend, San Diego, CA) for detection of live/dead cells. We used a BD FACSMelody cell sorter (BD Biosciences, San Jose, CA) to collect live cells. We labeled each group of NALT cells separately with the Sample-Tags from the BD Mouse Immune Single-Cell Multiplexing Kit (BD Biosciences, San Jose, CA), described in “Single Cell Labeling with the BD Single-Cell Multiplexing Kits” protocol. We then proceeded to library preparation with a mixture of ∼6000 cells (3000 cells from each group). We prepared the whole transcriptome using the BD Rhapsody System following the BD “mRNA Whole Transcriptome Analysis (WTA) and Sample Tag Library Preparation Protocol”. We assessed the quality and quantity of the final library by Agilent 4200 TapeStation system using the Agilent High Sensitivity D5000 ScreenTape (Agilent Technologies, Santa Clara, CA) and a Qubit Fluorometer using the Qubit dsDNA HS Assay, respectively. We diluted the final library to 3 nM concentration and used a HiSeq PE150 sequencer (Illumina, San Diego, CA) to perform the sequencing. scRNA-Seq
attributes.tag: GEO Accession
attributes.value: GSM4705884