title: GSM7675453: DN Cont2: MD37; Mus musculus; RNA-Seq
library_construction_protocol: Whole kidneys were harvested after cardiac perfusion with 0.9% saline, kidneys were finely minced in DMEM with 0.3mg/ml Liberase DL (Sigma/Roche) and homogenized at 37°C in MACS C-tubes using the gentle MACS dissociator (Miltenyi Biotec), digestion neutralized with PEB buffer (2 mM EDTA, 1% BSA in PBS), and cell suspension passed through 40μm cell strainer. Cells were then incubated with anti-mouse CD11b MACS beads (Miltenyi Biotech, 130-126-725) for 15 minutes, and CD11b+ cells separated by loading cells into MACS separator. The cell pellet was re-suspended in 350μl of buffer RLT (Qiagen), and RNA extracted with RNeasy Plus Mini Kits (Qiagen, 74134). RNA quality control was confirmed with RIN scores >7.9. cDNA library preparation was performed using a polyA-selected library preparation kit by the Vanderbilt Technologies for Advanced Genomics (VANTAGE) core facility