Action of circular RNAs on microbiome-gut-brain axis in mice

To find some more connections between circRNAs and gut microbiome using direct and indirect evidences.Mice were randomly allocated into two groups, control and model, 12 for each group. The mice of control group were fed with standard diet (Control), and the mice of the model groups were fed with high sugar & fat diet (Model), and water was available freely, these treatments were lasted 3 months.RNA sequencing libraries were prepared from 2μg of total RNA using the TruSeq Kit (Illumina, Inc.) with modifications. Instead of purifying poly-A RNA using poly-dT primer beads, we removed ribosomal RNA using the Ribo-Zero rRNA Removal Kit (Illumina, Inc.). Linear RNA was removed using RNase R (Epicentre, Inc). Poly-T oligo-attached magnetic beads (Illumina, Inc.) were used to remove residual poly-A RNA. All other steps were performed according to the manufacturer's protocols.