GSM8830299: MLN, naïve, vaccination, rep1; Mus musculus; RNA-Seq
Single cells were isolated from the lymph node by mechanical disruption through a 70 um cell strainer. Cells were washed once with RPMI+10%FBS and resuspended in PBS+0.04%BSA before filtering through a 35 um cell strainer. Cells were washed once with PBS+0.04%BSA before cell counting. Single cell suspensions were run on a Chromium X instrument and libraries were prepared following the Chromium Next GEM Single Cell 3' RNA-Seq - Dual Index Assay version 3.1 (10x Genomics, user guide CG000315, RevE) by the Cornell BRC Genomics Facility (RRID:SCR_021727). We targeted 10,000 cells and used 13 cycles of cDNA amplification. Sample quality was confirmed using a Qubit (DNA HS kit; Thermo Fisher) to determine concentration and a Fragment Analyzer (Agilent) to confirm fragment size integrity. Libraries were sequenced on a NovaSeqX, 10B flowcell with 2 x 150bp read length.