title: GSM4586763: brachial LN; Mus musculus; RNA-Seq
library_construction_protocol: Brachial, inguinal and mesenteric lymph nodes were harvested from Islet-CRE:Rosa-EYFP P2 pups into RPMI (Fisher Scientific) with 2% FBS (EuroClone), 10mM HEPES (Fisher Scientific). Tissue were transferred to room temperature digestion buffer (0,45mg/ml LiberaseTM (Roche) in FBS) and incubated at 37oC for 10 minutes at 900rpm in thermomixer. Tissue was pipetted 30 times with P1000 and incubated another 10 minutes. The digested cell suspension was resuspended in complete RPMI medium and centrifugated, then cells were resuspended in MACS buffer with anti-CD45 MACS microbeads (Miltenyi) to deplete CD45+ cells. The remaining cell suspension was spun down and immediately run on the 10X Chromium (10X Genomics) and then processed through library preparation following the recommended protocol for the Chromium Single Cell 3' Reagent Kit (v2 Chemistry, 10X Genomics). Libraries were run on the ……… Chromium Single Cell 3' Reagent Kit (v2 Chemistry, 10X Genomics)