GSM9216851: snRNA, tumor 10, kidney; Homo sapiens; RNA-Seq
From fresh frozen tissues, nuclei isolation was performed using low-retention microcentrifuge tubes throughout the procedure to minimize nuclei loss. Briefly, OCT-embedded tissue fragments were rapidly thawed on wet ice to remove excess OCT, mechanically dissociated by pipetting, and homogenized in TST solution. The homogenate was filtered through a 30 µm MACS SmartStrainer and pelleted by centrifugation for 4 minutes at 500 × g at 4°C. The resulting nuclei pellet was resuspended in 150 µL of 10x Genomics Diluted Nuclei Buffer. Trypan blue-stained nuclei were counted manually using INCYTO C-Chip Neubauer Improved Disposable Hemacytometers. Library construction according to the manufacturer's protocol (Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 User Guide, Rev D or Chromium Next GEM Single Cell Multiome ATAC + Gene Expression User Guide, Rev F). Libraries were normalized and pooled for sequencing on a NovaSeq SP-100 flow cell (Illumina, Inc., San Diego, CA, USA).