GSM5732075: KM, Lym, Prog, 5; Danio rerio; RNA-Seq
Adult and juvenile zebrafish were euthanized by rapid chilling and confirmed deceased prior to dissecting thymi and/or kidney marrows. Live cells were sorted (85 um nozzle) on a BD FACSAriaII into 300 μL of 0.5% BSA in DPBS and kept on ice following the sort. Between 50,000 and 100,000 cells were sorted per sample, assessed by Trypan blue staining, and then pelleted by centrifugation at 400g x 5 minutes and resuspended to a final concentration of 1000 cells/uL. Transcriptome profiling was performed using the Chromium Next GEM Single Cell 3' v3.1 Reagent Kits (10x Genomics, Inc.). A targeted recovery of 6,000 cells was set for loading the Chromium Controller in all assays and steps were performed as described in the Chromium Next GEM Single Cell 3' Reagent Kits v3.1 User Guide (Rev D). The final transcriptome libraries were pooled in sets of 4 or 5 and sequenced at the Harvard University Bauer Core (https://bauercore.fas.harvard.edu/) on an Illumina NovaSeq 6000 using an SP flow cell (Read 1: 28 cycles, i7 index: 8 cycles, i5 index: 0 cycles, Read 2: 91 cycles). scRNA-Seq
GEO Accession
GSM5732075