title: GSM7115127: AD1; Homo sapiens; RNA-Seq
library_construction_protocol: Tissue samples were processed as follows: First, they were washed with phosphate-buffered saline (PBS) and then minced into small pieces (approximately 1mm3) on ice. Next, they were enzymatically digested using 285 U/mL collagenase I (Worthington), 355 U/mL collagenase II (Worthington), 0.5 U/mL Dispase II (Sigma), and 50 U/mL DNase I (Worthington) for 45 min at 37°C with agitation. After digestion, the samples were sieved through a 70µm cell strainer and then centrifuged at 300g for 5 min. Following the removal of the supernatant, the pelleted cells were suspended in red blood cell lysis buffer (Miltenyi Biotec) to lyse any remaining red blood cells. The cell pellets were then washed with PBS containing 0.04% BSA and re-suspended in the same buffer. The samples were then re-filtered through a 35μm cell strainer to ensure single-cell dissociation. The dissociated single cells were stained with AO/PI to assess cell viability using the Countstar Fluorescence Cell Analyzer. For the scRNA-Seq libraries, the 10X Genomics Chromium Controller Instrument and Chromium Single Cell 3' V3 Reagent Kits (10X Genomics, Pleasanton, CA) were used.