title: GSM9435951: Alzheimer's Disease, PTP1B +/+, rep2; Mus musculus; RNA-Seq
library_construction_protocol: FACS-based viability enrichment and single-cell library preparation were carried out at the Cold Spring Harbor Laboratory Single-Cell Biology Shared Resource. Prior to FACS, Single-cell suspensions were washed by centrifugation at 4°C, 600xg in a swinging bucket rotor, and resuspended in 1X PBS with 0.04% BSA, and stained with SytoxBlue dead cell stain (ThermoFisher #S34857). Viable, intact cells were sorted using a Sony SH800 sorter on a 70um chip into PBS/BSA, pelleted as above and resuspended at 1,000 cells/μL. Single-cell RNA sequencing was performed using 10X Genomics NextGEM v3.1 3' Gene Expression kits by the Cold Spring Harbor Laboratory Single-Cell Biology Shared Resource. Cells were loaded into a 10x Genomics Chromium X instrument and libraries prepared according to the manufacturer's instructions. Unique dual-indexed libraries were paired end sequenced using the Illumina NextSeq2000 sequencing system with 28x10x10x90bp reads to a mean depth of ~50,000 reads per cell.