title: GSM8650736: IL1r_KO_infected; Mus musculus; RNA-Seq
library_construction_protocol: Kidneys were transferred into RNAlater solution (Sigma Aldrich, R0901) and kept at 4 overnight. The next day, the RNAlater solution was removed, and the tissues were snap frozen in liquid nitrogen until processing. Processing of the tissue was done at the day of sequencing. In short, tissues were thawed, then cut into small pieces using a surgical blade. Next, the tissues were lysed using a Dounce homogenizer with Nuclei EZ lysis buffer (Sigma Aldrich NUC-101). The tissues were ground with a loose pestle 12-15 times, then another 12-15 times with a tight pestle. Homogenized tissue was incubated 5 minutes on ice, filtered using a 40um sieve and washed with Nuclei EZ lysis buffer. Nuclei were then pelleted at 500g for 5 minutes and washed once with 1%BSA containing RNAse inhibitor (New England BioLabs, 0.2U/ul). Nuclei were briefly vortexed and strained through a 30um strainer. The quality of the nuclei was assessed under the microscope. Library preparation and RNA sequencing was performed by using Drop-​seq platform from 10X Genomics. 10000 cells per sample were loaded in to the 10X Chromium controller and library preparation was performed according to the manufacturer's indications