Instrument: Illumina HiSeq 1500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Embryonic kidneys were micro-dissected using Dumont #5 INOX surgical forceps in cold phosphate-buffered saline (PBS) pH 7.4 under a Leica EZ4D microscope and were washed in cold PBS immediately after microdissection. Kidney explants were dissociated using 0.3% Collagenase (Sigma Aldrich) for 10 minutes at 37°C. Kidneys were then triturated through a 25-gauge needle to make a single-cell suspension, and then washed 3 times in cold PBS supplemented with 2% fetal bovine serum. Before sorting, cells were run through a 40μm strainer (ThermoFisher Scientific). Cells were sorted into PBS (if undergoing second sort) or directly into Qiagen's RLT Lysis Buffer. Sequencing of samples was performed using the Illumina HiSeq system (Illumina, San Diego).