GSM8554268: control; Homo sapiens; RNA-Seq
Frozen tissue samples were prepared for single nuclei RNA sequencing using the Chromium Nuclei Isolation Kit (PN-1000493 10x Genomics). In brief, frozen tissue samples were dissociated in lysis buffer with a sterile plastic pestle. During the dissociation samples were stored on ice. Dissociated tissue was pipette mixed and incubated for 10 minutes on ice. Dissociated tissue was passed through a 100 µm filter prior to transferring to a nuclei isolation column. Samples were centrifuged and the flowthrough which contained nuclei was collected. Nuclei were washed and resuspended in 25-50 µL. Nuclei were counted using a hemocytometer. Following manufacturers protocols, nuclei were prepared for single nuclei RNA sequencing using the Chromium GEM-X single cell 3' Kit (10X Genomics).