title: GSM9191678: mpegGFP_MTZ_0h; Danio rerio; RNA-Seq
library_construction_protocol: 600 mpeg1:EGFP;myo6b:mCherry-NTR2.0;she:H2A-mCherry triple-positive zebrafish larvae were dissociated on ice using 0.25% Trypsin-EDTA through gentle pipetting for 8min. The dissociated tissue was then filtered through a 70 µm filter. Following several washing steps with ice-cold 1XDPBS/0.04% BSA, the cells were filtered again through a 70 µm strainer. To stain nucleated cells, the suspension was incubated with DRAQ5 for 5 min on ice. Cells were sorted by the Stowers Institute Cytometry Core Facility using BD Influx Cell Sorter (BD Biosciences) and BD FACSymphony S6 (BD Biosciences) instruments. Sorted cells were spun down at 350 rcf at 4 °C for 2 min in a swinging bucket centrifuge with slow deceleration. The cell pellet was gently resuspended in 100 µL of 1X DPBS/0.04% BSA. Cells with a viability of 80% or higher were loaded onto a Chromium X Series instrument (10x Genomics) and libraries prepared using the Chromium Next GEM Single Cell 3' Reagent Kit v3.1 (10x Genomics) according to manufacturer's instructions.
identifiers.namespace: GEO
identifiers.id: GSM9191678_r1