title: GSM4774865: ISC young rep2; Drosophila melanogaster; RNA-Seq
library_construction_protocol: Whole midguts were dissected in 1xPBS, 1% BSA and dissociated at RT using 0.5% Trypsin-EDTA solution. Midguts were agitated by vortexing every 30 min and dissociated cells were removed from the enzyme solution and placed on ice in 1xPBS, 1% BSA and 2%FBS every 30 min. Once the tissue was completely dissociated, cells were FACS sorted to ensure single cells were obtained after which cell viability was measured using a Vi-CELL analyzer to confirm >80% viability. The cells were then immediately applied to the 10XGenomics chromium platform. Sample processing for single-cell RNA-seq was done using Chromium Single Cell 3' Library and Gel bead kit v2 following manufacturer's user guide (10x Genomics).