The bacterial 16S V3 hypervariable region was amplified from provided gDNA template using the primerswith partial Illumina adapter (341F:TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTACGGGNGGCWGCAG and518R: GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGATTACCGCGGCTGCTGG)(Klindworth et al. 2013). PCRwas performed using SolarBio PCR Mastermix (SolarBio, China) using the PCR profile of : 95C for 2minutes followed by 30 cycles of 95C for 30 s, 50C for 15s and 72C for 15s. PCR products were purifiedusing SPRI Bead (Oberacker et al. 2019) followed by index PCR to incorporate Illumina dual indexbarcode. The barcoded libraries were inspected on gel, pooled according to band intensity and purifiedusing SPRI Bead. Quantification of the pooled libraries used Denovix high sensitivity and appropriateamounts of the libraries were loaded onto a NovaSEQ6000 (Illumina, San Diego) for 2 x 150 paired-endsequencing.