title: GSM5988094: GH4:318GCecum; Mus musculus; OTHER
library_construction_protocol: mRNA-seq: Total RNA was isolated from amygdala, hypothalamus or liver using TRI Reagent (MRC molecular research center, OH, USA). Total RNA from the liver was further purified using SimplyRNA tissue kit on Maxwell RNA extraction system (Promega). RNA quality was accessed using RNA 6000 Nano Series II Kit on Agilent 2100 Bioanalyzer (Agilent141 Technologies, Santa Clara, CA, USA). RNA integrity number for each tissue RNA was higher than 8.5. mRNA-seq: 200ng of Total RNA was constructed in a library using Smarter Stranded Total RNA Sample Prep Kit HI Mammalian (Clontech Laboratory, Inc.). The library was quantified using High Sensitivity DNA kit on Agilent 2100 Bioanalyzer and subjected to 50 bp single end sequencing analysis (55.7 million / sample on average) on a Hiseq system (Illumina). 16S DNA-seq: DNA was extracted from the cecal samples using the bead-beating method described by Odamaki et al (Odamaki T, Kato K, Sugahara H, Hashikura N, Takahashi S, Xiao JZ et al. (2016). Age-related changes in gut microbiota composition from newborn to centenarian: a cross-sectional study. BMC Microbiol. 16, 90.). 16S DNA-seq:  The 16S rRNA V3–V4 region was amplified using Tru357F (5'-CGCTCTTCCGATCTCTG TACGGRAGGCAGCAG-3') and Tru806R (5'-CGCTCTTCCGATCTGAC GGACTACHVGGGTWTCTAAT-3') primers and sequenced on Illumina MiSeq.