DNA-Seq of feline gut microbiome
Fecal DNA samples were extracted using the Qiagen Allprep PowerFecal DNA/RNA kit (Qiagen, Redwood City, CA, USA). The homogenization step was conducted using the Qiagen PowerLyzer24 instrument. An aliquot of 500 ng DNA from each sample was sheared into 500-bp fragments using an M220 Focused-ultrasonicator (Covaris, Woburn, MA, USA). The WGS metagenomic libraries were prepared using NEBNext Ultra II DNA Library Prep Kit for Illumina (New England BioLabs, Ipswich, MA, USA). TapeStation 4200 (AgilentAgilent Technologies, Santa Clara, CA, USA) was utilized to assess the library size distributions. Subsequently, the final libraries were quantified using qPCR prior to sequencing on an Illumina NovaSeq6000 sequencing machine, employing the 150-bp paired-end mode, by Novogene (Novogene Corporation Inc., Sacramento, CA, USA).