GSM8194524: P479, Day 5 GFP, rep 1; Homo sapiens; RNA-Seq
Islets were briefly washed in PBS and dispersed into single cells. Cells were sorted using a FACS Jazz or FACS Aria III (BD Biosciences, Franklin Lakes, NJ, USA). Live single cells were selected based on DAPI exclusion and FSC, sorted into 384-well hard shell plates (BioRad, Hercules, CA, USA) and further processed with SORTseq (SOrting and Robot-assisted Transcriptome sequencing) as previously described (Muraro, et al 2016). Mapping of the sequenced reads and quantification of the number of transcripts reads 2 from the fastq files were mapped to the human reference transcriptome with BWA and reads 1 were used to extract the cell-barcode sequences and unique molecular identifiers (UMI). UMI-correction was performed to the read and barcode counts resulting in unique transcript count tables. Spike-in reads were removed from the dataset.