GSE117590
Mouse gonadal transcriptome at 12.5dpc, 13.5dpc, 16.5dpc and 6dpp.
we employed RNA-Seq to examine transcriptome profiles of male and female mouse gonads at 12.5dpc, 13.5dpc, 16.5dpc and 6dpp. Methods: Gonadal mRNA profiles of 12.5dpc,13.5dpc, 16.5dpc and 6dpp mice were generated by deep sequencing, in triplicate, using Illumina Hiseq 2500. The cDNA library was constructed with a SMARTer® Ultra Low Input RNA for lllumina® Sequencing kit (Clontech Laboratories) and sequenced on an Illumina HiSeq 2500.After sequencing, clean reads were obtained by removing reads containing the adaptor sequences, reads with > 5% ambiguous bases, and low-quality reads, then mapped to the mouse genome (version: mm10_GRCm38) using TopHat software. Gene expression level was calculated using the fragments per kilobase per million mapped reads method. Overall design: RNA sequencing was employed to examine transcriptome profiles of murine 4 female and 4 male gonads at developmental stages involving in sex differentiation, meiosis, and gametogenesis . Analysis of Differentially Expressed Genes (DEGs) was performed by the DESeq package in R language. Wayne analysis was then performed to screen sex-biased expressed genes. Genes were submitted to the databases of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) for enrichment analysis of the significant GO terms and KEGG pathways. The Series-Cluster analysis of expression profiles of DEGs was performed by using the STEM method. Weighted gene co-expression network analysis (WGCNA) was used to create a co-expression network. Alternatively spliced transcripts were identified by the software AS detector (ASD) using the Fisher's exact test.