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In short, a targeted PCR-based sequencing approach was used, where the V3V4 regions of the 16S rRNA gene were targeted to generate amplicons using the primers 341F (5-CCTAYGGGRBGCASCAG-3) and 806R (5-GGACTACNNGGGTATCTAAT-3) with the barcode

By heating water to its boiling point (100C or 212F), all bacteria and other microorganisms are killed

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