GSM2422366: C_TH_21; Mus musculus; RNA-Seq
The medial prefrontal cortex was dissected, minced, and resuspended in HBSS. 1% of the suspension was removed and the remaining cells were manually dissociated in Neural Tissue Dissociation Buffer (P) (Miltenyi Biotec). Myelin was removed using a 30% percoll gradient and microglia were captured using Cd11b microbeads (Miltenyi Biotec). Cells were spun at 1,000 x g and flash frozen in liquid nitrogen. RNA was harvested using Qiagen Rneasy Micro Kit (Cat No. 74004). Samples were Poly(A) selected using the Poly(A) Purist Kit (Thermo Fisher Scientific Inc.) and prepared using NEBNext® Ultra™ Directional RNA Library Preparation Kit (New England BioLabs, Ipswich, MA). Samples went through 15 cycles of PCR. Samples were sequenced on the NextSeq 500 (Illumina, San Diego, CA) using (75 bp; paired-end reads) to a minimum target depth of 20 million reads per sample.
GEO Accession
GSM2422366