GSM3665814: 4 Week-Cort-5; Mus musculus; RNA-Seq
Brains were coronally cryosectioned onto RNAse free slides. Sections were fixed and rehydrated in ethanol, stained with cresyl violet, and dehydrated in a graded ethanol series. The entire hypothalamic arcuate nucleus was dissected from each sample using laser capture microdissection. RNA was extracted using an RNeasy Micro Plus Kit (Qiagen) according to manufacturer's instructions and analyzed for RNA integrity on an Agilent 2100 Bioanalyzer using an RNA 6000 Pico Kit. All RNA that was subsequently used had RNA integrity (RIN) scores >6.5. RNA was converted to cDNA using the Ovation RNA-seq System V2 (NuGEN). The resultant cDNA was then fragmented to an average size of ~200bp using Bioruptor Pico (Diagenode) before end repair and adaptor ligation was performed using the Ovation Rapid DR Multiplex System 9-16 (NuGEN) to allow multiplex sequencing. Prepared libraries were quantified using KAPA Library Quantification Kit (KAPA Biosystems) before sequencing in a single lane on a HiSeq4000 (Illumina; 2 day samples) or HiSeq2500 (Illumina; 4 week samples).
GEO
GSM3665814