title: GSM4648636: Control-1-2; Rattus norvegicus; RNA-Seq
library_construction_protocol: Animals were rapidly decapitated 18~19 h after the worker-parasite relationship transpired in group testing. Additional four animals not exposed to the Skinner box were sacrificed as home-cage controls. Brains were removed and coronal slices (2 mm) were fleshly dissected in a cold brain matrix. ACC were then microdissected, using a dissecting microscope and reference coordinates obtained from the Paxinos and Watson rat brain atlas. RNA was isolated with TriZol reagent (Invitrogen) and purified with RNeasy Micro Kits (Qiagen). All RNA samples were determined to have RIN values > 8.5 (BioAnalyzer, Agilent) for RNA-sequencing. RNA quality was assessed by analysis of rRNA band integrity on an Agilent RNA 6000 Nano  kit (Agilent Technologies, CA). Ahead of cDNA library construction, the 1ug of total RNA and magnetic beads with Oligo (dT) were used to enrich poly (A) mRNA from it. Then, the purified mRNAs were disrupted into short fragments, and the double-stranded cDNAs were immediately synthesized. The cDNAs was subjected to end-repair, poly (A) addition, and connected with sequencing adapters using the TruSeq RNA sample prep Kit (Illumina, CA). The suitable fragments automatically purified by BluePippin 2% agarose gel cassette (Sage Science, MA) were selected as templates for PCR amplification. The final library sizes and qualities were evaluated electrophoretically with an Agilent High Sensitivity DNA kit (Agilent Technologies, CA) and the fragment was found to be between 350–450 bp. Subsequently, the library was sequenced using an Illumina NovaSeq sequencer (Illumina, CA).