16S rRNA metagene
The 16S rRNA genes' V3V4 hypervariable region was amplified using 341F forward primer containing IonCode barcode sequences provided by Ion Torrent protocol and 805R reverse primer. The amplicon library samples were pooled in equimolar concentration in the final library which was then used for template preparation in Ion Chef Instrument using Ion 520 & Ion 530 ExT KitChef and Ion 530 semiconductor chip. The prepared Ion 530 chip was loaded onto Ion Gene studio S5 for sequencing.