title: GSM9053411: 4s2 Global Cre, rep2; Mus musculus; RNA-Seq
library_construction_protocol: Mice were anesthetized via 5.0% isoflurane before exsanguination and transcardial perfusion with ice-cold Dulbecco's phosphate buffered saline (DPBS; Gibco cat. #14040133). Following perfusion, brains were quickly removed and whole right hemispheres sans brainstem and cerebellum were quickly minced using forceps on top of an ice-chilled petri dish. Minced tissue from the hemispheres (n=3 APOE4s2G, n=3 APOE4s2A, n=1 APOE4s2, n=2 E2-TR, and n=1 E4-TR) were immediately transferred into gentleMACS C-tube (Miltenyi cat. #130–093-237) containing Adult Brain Dissociation Kit (ADBK) enzymatic digest reagents (Miltenyi cat. #130–107-677) prepared according to manufacturer's protocol. Tissues were dissociated using the “37C_ABDK” protocol on the gentleMACS Octo Dissociator instrument (Miltenyi cat. #130–095-937) with heaters attached. After tissue digestion, cell suspensions were filtered through 70 μm mesh cell filters to remove debris following the manufacturer's suggested ABDK protocol. The resultant suspension was sequentially filtered (x2) using fresh 30 μm mesh filters. Cell viability was checked using Acridine Orange/Propidium Iodide (AO/PI) viability kit (Logos Biosystems cat. #LGBD10012). Following viability and counting, cells were diluted to achieve a concentration of ~ 1000 cells/100uL. The diluted cell suspensions were loaded onto the 10x Chromium. Each sample was loaded into a separate channel on the Single Cell 3′ Chip and libraries were prepared using the Chromium v3 Single Cell 3′ Library and Gel Bead Kit (10x Genomics). Final library quantification and quality check was performed using BioAnalyzer (Agilent).