GSM9177251: Alpha cells, E17.5 Gcg-Timer, green dominant and green/red copositive cells; Mus musculus; RNA-Seq
Pancreata from Gcg-Timer embryos and wild-type littermates were dissected at E17.5　and dissociated using 0.5 g/L trypsin/0.53 mmol/L-EDTA solution (Nacalai Tesque, Kyoto, Japan) at 37 °C for 5 minutes. The dissociated cells were resuspended in PBS containing 1% FBS, and newly generated and differentiated α cells were sorted using BD FACSAria™ III Cell Sorter (BD Biosciences, San Jose, CA, USA). Single-cell cDNA libraries were prepared using the 10× Genomics Chromium™ platform (Pleasanton, CA, USA). The Single Cell 3' Reagent Kits v2 were used to generate single-cell libraries. The libraries were pooled and sequenced using an Illumina NextSeq500 platform (San Diego, CA, USA) with a 150 cycle High Output v2 kit in paired-end format with 26 bp Read 1, 8 bp I5 Index, and 85 bp Read 2.