GSM8973926: bMs, wt, 3dpf, fli1a+ mpeg+, plate1; Danio rerio; RNA-Seq
Embryos were collected and anesthetized in E3 medium containing 0.01% tricaine and placed in ice-cold PBS. After centrifugation, the pellet was washed with ice-cold PBS and resuspended in TrypLE (ThermoFisher, 12563011) for 45 min at 32°C with vigorous shaking. Cells were then washed with FACS buffer (PBS + 2% FBS + 2mM EDTA), dissolved in FACS buffer supplemented with DAPI (ThermoFisher, 0.5 mg/ml final concentration) and subsequently filtered through 40-μm nylon mesh. Cells were sorted on a FACS Aria II (BD Biosciences) 384 plates were processed by Single Cell Discoveries using the SORT-seq protocol (Muraro, M. J. et al. A Single-Cell Transcriptome Atlas of the Human Pancreas. Cell Syst. 3, 385-394.e3 (2016)). Illumina sequencing libraries were prepared with the TruSeq small RNA primers (Illumina) and paired-end sequenced at 75 bp read length on the Illumina NextSeq platform.