GSM9108724: D4 Lymph Node Gene Expression; Mus musculus; RNA-Seq
Cell density was adjusted to analyze 10,000 cells per sample. Single-cell gene expression (scRNA-seq) was performed using Chromium Single Cell 5' Library and Gel Bead Kit with V(D)J enrichment kit (10x Genomics). Gel Bead-in-Emulsion reverse transcription (GEM-RT) reaction, clean-up, and PCR amplification were performed to obtain cDNA for library generation. Single cell sequencing libraries were generated using 5'GEXv1 according to manufacturer's protocol. Libraries were uniquely indexed for multiplexed sequencing. Libraries were single i7-indexed using the Chromium i7 Multiplex kit (indices listed in table). Following isolation and clean-up of library DNA, concentration and fragment size were assessed using an Agilent Bioanalyzer and quantification by Qubit analysis (ThermoFisher).