16S Seq of Felis catus: feces
Total DNA was extracted from frozen feces samples by using PowerFecal DNA isolation kit (MoBio, Carlsbad, CA) according to the manufacturers instructions with modification by introducing sonication step before placing the bead tubes with feces sample horizontally for vortexing 15 minutes.  PCR amplification was performed by using the primer pairs 341F and 806R spanning the V3-V4 hypervariable regions of 16s rRNA gene along with Illumina adapters (Liu et al 2012).  Amplicon sequencing was performed followed by Illuminas 16S metagenomic sequencing library preparation protocol (Part # 15044223 Rev. A). The sequences were de-multiplexed based on the dual index sequences by employing Miseq built-in metagenomics workflow to obtain FASTQ files. FASTQ sequence files were processed employing QIIME by using standard parameters (Caporaso et al. 2010). FASTQ files were obtained from the MiSeq instrument system in pairs, where each pair represents two sets of reads per biological sample. The files were pre-processed to be made into contigs from pairs of reads, chimeras removed.