Illumina HiSeq 2500 paired end sequencing; RNA-seq experiment in J20 hAPP mice and littermate controls measuring total RNA in lysate of microdissected dentate gyrus
RNA-seq experiment in J20 hAPP mice and littermate controls measuring total RNA in lysate of microdissected dentate gyrus
Hemibrains were frozen at -80, then embedded in agar and dentate-microdissected using a tissue chopper and microdissection scissors under a dissection microscope, in chilled RNAase-free PBS.  Hemibrains were frozen at -80, then embedded in agar and dentate-microdissected using a tissue chopper and microdissection scissors under a dissection microscope, in chilled RNAase-free PBS.  RNA extraction was performed using adapted instructions from the Qiagen RNeasy Mini kit (74106). Briefly, dentate samples were submerged in RLT/β-mercaptoethanol buffer, minced with small scissors, and homogenized by passing the lysate through a 21G needle 15 times. Samples were centrifuged and the supernatants were transferred to new tubes. RNA was then purified according to the kit instructions, and eluted with nuclease-free water. Final RNA concentration was determined using a NanoDrop One spectrophotometer.  Approximately 300 ng of RNA extracted from the dentate gyrus of APP mice with high ∆FosB expression and from wild-type NTG mice (4 per genotype) were submitted to the University of Pennsylvania Next-Generation Sequencing Core, where library preparation was performed using Illumina TruSeq Total kits.
Experimental Factor: genotype
wild type genotype