title: GSM6919116: Microglia, APP/PS1, p38+/+, animal 2; Mus musculus; RNA-Seq
library_construction_protocol: PFA-fixed left brain hemispheres were dounce homogenized. Microglial-containing fractions were separated by 30% Percoll gradient (GE #17-5445-01) in RPMI 1640 (Gibco #11835030). Cells were incubated with 1:100 Zombie NIR dye solution (Biolegend #423106), and 1:50 CD11b-VioB (Miltenyi Biotec Cat# 130-113-810, RRID:AB_2726327) and P2ry12-PE (BioLegend cat. No. 848004; RRID: AB_2721645) antibodies. Approximately 10,000 live cells co-expressing CD11b and P2ry12 were sorted per mouse using a Sony Cell Sorter and immediately lysed in RLT Plus buffer and RNA extracted by RNeasy Plus Micro Kit (Qiagen #74034) according to manufacturer's instructions. Samples with high RIN values (M = 8.3, minimum = 6.7, maximum = 9.1) were sent to Novogene for transcriptome sequencing. cDNA library was generated with NEBNext Ultra II RNA kit and run on Illumina Platform PE150