GSM7337029: P0 mouse, snATAC-seq, rep M2; Mus musculus; ATAC-seq
From the harvested P0 mouse kidneys, nuclei were isolated as previously described in(Wilson et al., 2019). Briefly, flash frozen kidney pieces were thawed on ice and dissected into smaller pieces (<1mm) with a razor blade and then dounced in cold Nuclei EZ Lysis Buffer (Sigma, N3408) supplemented with protease inhibitor (Roche; 05892791001) and RNAse inhibitors (Promega, N2615; Thermo Fisher Scientific, AM2696). The tissue was dounce grinded 30 times with a loose pestle (Sigma, P0485), and then filtered through a 200um strainer (pluriSelect, 43-50200). The tissue was dounce grinded 10 times with a tight pestle, and then incubated for 5 mins on ice. Cells were filtered through 40uM filter (pluriSelect ,43-50040) and pelleted at 500g for 5min at 4°C in a swinging bucket centrifuge. Supernatant was removed and nuclei pellet resuspended in Nuclei EZ lysis buffer with RNAse inhibitors, incubated an additional 5 mins on ice and pelleted again. The final pellet was resuspended in 4°C Nuclei Buffer (10X Genomics, #2000207). Chromium_NextGEM_SingleCell_ATAC_ReagentKits_v1.1_UserGuide (CG000209)