title: GSM8141818: C57BL/6 mouse without ligature 3 [C3]; Mus musculus; RNA-Seq
library_name: GSM8141818
library_construction_protocol: Mouse glomeruli were isolated from the kidneys of perfused mice with Dynabeads M-450 Tosylactivated in PBS. Briefly, the dissected kidneys (1 and 1/2 kidneys) from each mouse were minced and digested in 1 mg/ml of collagenase A (10103578001; Sigma-Aldrich, St-Louis, MO, USA) and 100 U/ml DNase Ⅰ in Hank's Balanced Sult Solution (14175095; Thermo Fisher Scientific, Waltham, MA, USA) at 37℃ for 30 minutes. Then, the digested mixture was passed through a 100 µm cell strainer (542000; Greiner, Kremsmünster, Australia) and placed on a magnet for purification. After washing with PBS twice, glomeruli were collected. The purity of glomeruli was confirmed more than 95% by microscopy. Glomerular RNA was isolated using PureLink RNA Mini Kit (12183018A; Invitrogen) according to the manufacturer's guidance. A total amount of 1 μg RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using NEBNext® UltraTM RNA Library Prep Kit for Illumina® (NEB, USA) following manufacturer's recommendations and index codes were added to attribute sequences to each sample.