GSM9059712: Low, BMI 24.6, Age 58; Homo sapiens; RNA-Seq
The Chromium Nuclei Isolation Kit (10x Genomics, PN-1000493) was used to isolate single nuclei. Tissue samples were thawed and dissociation tubes (PN-2000564) pre-chilled on dry ice. Samples were dissociated on wet ice using lysis buffer and pestle agitation until samples reached homogenization. Dissociated tissues were centrifuged in nuclei isolation columns (PN-2000562) for initial removal of cell debris. To remove remaining cell debris, pelleted nuclei were washed by resuspending nuclei pellet once in Debris Removal Buffer and three times in Wash and Resuspension Buffer respectively, with centrifugation between washes. Nuclei were resuspended in Wash and Resuspension buffer on ice for further analysis. RNA-seq was performed by the Wayne State University Genome Sciences Core. After sequencing, the snRNA-seq library was demultiplexed using CellRanger mkfastq. Then, the CellRanger count was used to align the reference genome and tabulate counts by gene region.