Illumina HiSeq 2500 sequencing; GSM3573756: CER_405_Virgin; Mus musculus; RNA-Seq

CER_405_Virgin

RNA-seq experiment on five mouse brain regions-- --comparing expression in virgin females to primiparous females three weeks after pup weaning. For each brain region there are 8 replicates, though three samples were not used in the final analysis based on inspection of PCA plots (1 sample from the Hippocampus, Cerebellum and Amygdala data sets; see details below). Overall design: Comparison of gene expression in five brain regions between eight virgin and eight primiparous females three weeks after pup weaning. Pair-wise comparisons were performed using EdgeR transformed data and Limma Voom for statistical analysis.

RNA-seq experiment on five mouse brain regions--hypothalamus, hippocampus, neocortex, cerebellum, and the amygdala --comparing expression in virgin females to primiparous females three weeks after pup weaning.

Submitted on 02-MAR-2019

Brain samples were harvested on the same day and snap frozen in RNA later solution (Ambion), within a two-hour window. Frozen brain tissue was shipped to FSU.RNA and protein were extracted from the frozen brain tissue using the NucleoSpin RNA/Protein purification kit (Macherey-Nagel). The brain tissue was homogenized using a motorized homogenizer 1 ug of total RNA was used for subsequent mRNA isolation (New England Biolabs Inc. PolyA magnetic mRNA isolation kit). The External RNA Controls Consortium (ERCC) RNA Spike-in Controls (Ambion 4456740) were diluted 1:100 and added to total RNA before mRNA isolation. RNA-seq libraries were generated using the NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs Inc. E7530L). The qualities of the libraries were examined using Agilent High Sensitivity DNA Bioanalyzer Chips (Agilent Technologies 5067-4626) and quantified by KAPA Library Quantification Kits Libraries were sequenced on an Illumina 2500 platform, single end, 100 base reads.

RNA-seq experiment on five mouse brain regions--hypothalamus, hippocampus, neocortex, cerebellum, and the amygdala --comparing expression in virgin females to primiparous females three weeks after pup weaning.

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