GSM6434044: Fgf9 Null; Mus musculus; RNA-Seq
Pancreata were minced into small pieces and dissociated into single cells using TrypLE Express dissociation reagent (Gibco, 12604013) at 37°C for 15 min. We neutralized dissociations with FACS buffer (10% FBS+ 2 mM EDTA in phenol-red free HBSS), and passed cell suspensions through 30 µm cell strainers. Single cells were stained with DAPI before loading to the sorter (BD FACSAria II). After size selection to remove doublets, all DAPI negative live cells were collected. Libraries were prepared according to the manufacturer's instructions (10x Genomics). Each resulting library was sequenced on the Novaseq 6000 platform (Illumina) using the S4 flowcell with the following parameters: Read 1 – 28 cycles, Index 1 i7 – 8 cycles, Index 2 i5 – 0 cycles, Read 2 – 91 cycles.