Illumina HiSeq 2500 paired end sequencing; GSM3145861: ovary_CV1; Mus musculus; RNA-Seq

ovary_CV1

Gut microbiota and the circadian clock are both key regulators of the metabolic processes. Although recent evidence points to the impact of the circadian clock on microbiota, gut microbiota effect on diurnal host gene expression remains elusive. A transcriptome analysis of germ-free mice reveals subtle changes in circadian clock gene expression. However, a lack of microbiome leads to liver feminization and alters the expression of male-specific genes involved in lipid metabolism and xenobiotic detoxification associated with sustained activation of the Growth Hormone pathway. These results emphasize the mutual interaction of gut microbiota and its host even on unexpected functions. Overall design: Total RNA-Seq of testis and ovaries of conventional raised (convR) and Germ-free (GF) female mice under ad libitum feeding regime.

Total RNA-Seq of testis and ovaries of conventional raised (convR) and Germ-free (GF) female mice under ad libitum feeding regimen.

Submitted on 02-SEP-2018

Ovaries and testis were disrupted and extracted using the tissue lyser II (Qiagen) and the miRNeasy Mini Kit (Qiagen) following the manufacturer's protocol. The RNA were quantified with Ribogreen (Life Technologies) and quality assessed on a Fragment Analyzer (Advances Analytical). Sequencing libraries were prepared from 250 ng total RNA using the TruSeq Stranded Total LT Sample Prep Kit (Illumina) with the Ribo-Zero Gold depletion set. The procedure was automated on a Sciclone NGS Workstation (Perkin Elmer). The manufacturer's protocol was strictly followed, except for the PCR amplification step. The latter was run for 12 cycles with the KAPA HiFi HotStart ReadyMix (Kapa BioSystems). This optimal PCR cycle number has been evaluated using the Cycler Correction Factor method as described previously (Atger et al., PNAS, 2015). Purified libraries were quantified with Picogreen (Life Technologies) and the size pattern was controlled with the DNA High Sensitivity Reagent kit on a LabChip GX (Perkin Elmer). Libraries were then pooled by 24 and each pool was clustered at a concentration of 7 pmol on 8 lanes of a v3 paired-end sequencing flow cell (Illumina). Sequencing was performed for 2 x 100 cycles on a HiSeq 2500 strictly following Illumina's recommendations.

Total RNA-Seq of testis and ovaries of conventional raised (convR) and Germ-free (GF) female mice under ad libitum feeding regimen.

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