Method of Microglial DNA-RNA Purification from a Single Brain of an Adult Mouse

Microglia, the resident brain immune effectors cells, show dynamic activation level changes for most neuropsychiatric diseases, reflecting their complex regulatory function and potential as a therapeutic target. Emerging single-cell molecular biology studies are used to investigate the genetic modification of individual cells to better understand complex gene regulatory pathways. Although multiple protocols for microglia isolation from adult mice are available, it is always challenging to get sufficient purified microglia from a single brain for simultaneous DNA and RNA extraction for subsequent downstream analysis. Moreover, for data comparison between treated and untreated groups, standardized cell isolation techniques are essential to decrease variability. Here, we present a combined method of microglia isolation from a single adult mouse brain, using a magnetic bead-based column separation technique, and a column-based extraction of purified DNA-RNA from the isolated microglia for downstream application. Our current method provides step-by-step instructions accompanied by visual explanations of important steps for isolating DNA-RNA simultaneously from a highly purified microglia population.

Automated summary

Microglia, the resident immune cells in the brain, show dynamic activation level changes in neuropsychiatric diseases, making them a potential therapeutic target. Investigating genetic modifications of individual cells through single-cell molecular biology studies can help understand complex gene regulatory pathways. Isolating sufficient purified microglia from a single brain for DNA and RNA extraction remains challenging, and standardized cell isolation techniques are essential for data comparison.