GSM3454937: SAM24306366 SST; Mus musculus; RNA-Seq
Isolation of pure populations of neurons from adult mouse cortex using fluorescence activated cell sorting (FACS) was performed following published protocols 56 with the following modifications:  After enzymatic digestion with papain and trituration to obtain a homogenous cell suspension, cells were washed with 5ml of Hibernate A- LF twice at 1800 x g and resuspended in 5 ml of Hibernate A with 2.5ul of DAPI (1 ug/ml stock) added for FACS sorting. Neurons were sorted as DAPI(-):GFP(+) or DAPI(-):tdTomato(+) (depending on the transgenic reporter line) using a 100 micron filter in the ARIA sorter. Sorted cells (collected live) in Hibernate A medium were spun at 2000 x g at 4deg C for 5 min and cell pellets were processed for RNA extraction using a Qiagen RNeasy Plus Micro kit. RNA quality was checked using BioAnalzyer and cell type purity was checked using multiple TaqMan markers specific to the cell type of interest using BioMark Fluidigm chips. 500 ng RNA was submitted for sequencing after the samples passed these quality checks.
GEO
GSM3454937