title: GSM5098672: X_21; Homo sapiens; RNA-Seq
library_construction_protocol: Bulk RNA-seq: Muscle biopsies were homogenized using a mortar and pestle with liquid nitrogen. RNA was extracted from the powdered samples using the RNeasy Fibrous Tissue Mini Kit (Qiagen) and the QIAcube automatic processor (Qiagen). Integrity and concentration of the RNA were assessed using the Tapestation 4200 (Agilent Technologies), with the cut-off for acceptable integrity being an RNA integrity number (RIN) >7. Batch-tag-seq libraries were then produced from the RNA and run on the HiSeq 4000 by the DNA Technologies Core at U.C. Davis. Single-nuclei RNA-seq: Nuclei were isolated from the biopsies using the Singulator instrument (S2 Genomics). The instrument was primed with cold nuclei isolation and storage buffers (S2 Genomics) and the biopsy was loaded into the cartridge and covered with the 19.7 mm grinding cap. The “Nuclei_All_Tissues” protocol was used to isolate nuclei, after which the nuclei were centrifuged for 5 minutes and resuspended in buffer. The nuclei were counted using a Countess II Automated Cell Counter (Invitrogen), centrifuged again for 5 minutes and resuspended at the proper concentration for use in the Chromium Single Cell 5' Library and Gel Bead Kit v1 (10X Genomics). Samples were processed using the Chromium Single Cell A Chip Kit and Chromium Controller (10X Genomics). Quality control was performed on the Tapestation 4200 (Agilent Technologies). Libraries were sequenced in one lane of a NovaSeqS4 by the U.C. Davis Genomics Core.