metabarcoding of Felis catus
The hypervariable V4 region of the 16S ribosomal RNA gene was sequenced using Illumina technology, specifically the NovaSeq 6000 platform, which produced paired end reads of 150 base pairs (bp). To prepare the samples, a minimum DNA quantity of 200 ng was required, along with a purity ratio between 1.8 and 2.0 (260/280 ratio), a volume greater than 40 l, and a concentration exceeding 5 ng/l. The primers used for amplifying the V4 region were as follows: 515F: GTG CCA GCM GCC GCG GTA A and 806R: GGA CTA CHV GGG TWT CTA AT. Fragment selection was performed to obtain fragments approximately 300 bp in length.