GSM7401907: 7_Tg26_C16_B; Mus musculus; RNA-Seq
Nuclei from frozen mouse kidney outer cortex tissue samples and glomeruli-enriched samples were prepared following the protocol. Briefly, ~ 8 mm3 tissue fragments were cut by razor blade in EZlysis buffer (#NUC101-1KT, Sigma, Darmstadt, Germany) and homogenized 30 times using a loose Dounce homogenizer and 5 times by tight pestle. After 5 mins of incubation, the homogenate was passed through a 40 µm filter (PluriSelect, El Cajon, CA) and centrifuged at 500g at 4°C and for 5 min. The pellet was washed with EZlysis buffer (Sigma Aldrich, St Louis, MO) and again centrifuged at 500g at 4°C for 5 min. The pellet was resuspended with DPBS with 1% FBS and passed through a 5 µm filter (PluriSelect, El Cajon, CA) to make final nuclei prep for loading on to a 10xChromium Chip G (10X Genetics, San Francisco, CA) for formation of gel beads in emulsion (GEM). 10x Genomics Chromium 3' V3.1 mRNA-seq library protocol