title: GSM7790650: M-KO-PFF-Microglia; Mus musculus; RNA-Seq
library_construction_protocol: Once sorted, microglia (200000-500000 cells) and macrophage (10000-30000 cells) cell suspensions (>90% purity after sorting) were centrifuged at 4°C and 400xg for 5 minutes, resuspended in RLT Plus Lysis buffer (QIAGEN, Germany) with 1% β-Mercaptoethanol and vortexed for 1 minute for cell lysis. The solution was further homogenized using a 1mL syringe and a 21gag needle. Total RNA was extracted using RNeasy® Mini Kit (QIAGEN) according to the manufacturer's protocol. Total purified RNA was sent to BGI Hong Kong where Switching Mechanism at 5' End of RNA Template (SMART-seq2) sequencing service was requested. Samples were tested for quality control using Agilent 2100 Bio analyzer (Agilent RNA 6000 Nano Kit). RNA integrity (RNI) >7 was considered suitable for sequencing. Gene expression analysis was performed through full population RNA sequencing using the SMART-seq2 method according to BGI's standard methodology.