SRX3427328 T2.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718837 SAMN08040949 T2.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427329 T2.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718837 SAMN08040949 T2.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427330 T1.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718838 SAMN08040948 T1.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427331 T1.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718838 SAMN08040948 T1.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427332 T1.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718838 SAMN08040948 T1.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427333 T0.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718839 SAMN08040947 T0.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427334 T0.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718839 SAMN08040947 T0.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427335 T0.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718839 SAMN08040947 T0.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427336 T16.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718841 SAMN08040963 T16.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427337 T3.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718840 SAMN08040950 T3.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427338 T2.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718837 SAMN08040949 T2.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427339 T19.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718842 SAMN08040966 T19.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427340 T18.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718843 SAMN08040965 T18.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427341 T18.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718843 SAMN08040965 T18.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427342 T18.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718843 SAMN08040965 T18.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427343 T17.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718844 SAMN08040964 T17.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427344 T17.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718844 SAMN08040964 T17.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427345 T17.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718844 SAMN08040964 T17.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427346 T16.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718841 SAMN08040963 T16.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427347 T19.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718842 SAMN08040966 T19.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427348 T19.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718842 SAMN08040966 T19.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427349 T4.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718845 SAMN08040951 T4.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427350 T4.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718845 SAMN08040951 T4.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427351 T3.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718840 SAMN08040950 T3.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427352 T3.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718840 SAMN08040950 T3.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427353 T5.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718846 SAMN08040952 T5.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427354 T5.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718846 SAMN08040952 T5.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427355 T5.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718846 SAMN08040952 T5.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427356 T4.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718845 SAMN08040951 T4.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427357 T6.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718847 SAMN08040953 T6.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427358 T6.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718847 SAMN08040953 T6.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427359 T13.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718848 SAMN08040960 T13.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427360 T12.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718849 SAMN08040959 T12.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427361 T11.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718850 SAMN08040958 T11.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427362 T11.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718850 SAMN08040958 T11.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427363 T11.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718850 SAMN08040958 T11.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427364 T12.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718849 SAMN08040959 T12.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427365 T13.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718848 SAMN08040960 T13.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427366 T13.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718848 SAMN08040960 T13.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427367 T14.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718851 SAMN08040961 T14.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427368 T12.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718849 SAMN08040959 T12.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427369 T14.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718851 SAMN08040961 T14.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427370 T15.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718852 SAMN08040962 T15.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427371 T15.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718852 SAMN08040962 T15.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427372 T15.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718852 SAMN08040962 T15.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427373 T16.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718841 SAMN08040963 T16.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427374 T10.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718853 SAMN08040957 T10.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427375 T20.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718854 SAMN08040967 T20.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427376 T20.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718854 SAMN08040967 T20.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427377 T10.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718853 SAMN08040957 T10.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427378 T8.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718855 SAMN08040955 T8.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427379 T9.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718856 SAMN08040956 T9.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427380 T8.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718855 SAMN08040955 T8.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427381 T8.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718855 SAMN08040955 T8.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427382 T7.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718857 SAMN08040954 T7.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427383 T7.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718857 SAMN08040954 T7.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427384 T6.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718847 SAMN08040953 T6.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427385 T7.1 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718857 SAMN08040954 T7.1 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427386 T10.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718853 SAMN08040957 T10.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427387 T9.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718856 SAMN08040956 T9.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427388 T9.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718856 SAMN08040956 T9.3 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427389 T14.2 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718851 SAMN08040961 T14.2 AMPLICON METAGENOMIC PCR Illumina MiSeq SRX3427390 T20.3 SRP125772 PRJNA419221 Amplicon libraries of V6ÐV8 regions of 16S rRNA genes were obtained using primers 341F (5Õ-CCTACGGGIGGCWGCAG-3Õ) and 805R (5Õ-GACTACHVGGGTATCTAATCC-3Õ) (Al Kharusi et al. 2016) containing the Illumina-specific MID barcodes. The PCR mixture contained 10 pM of each primer, 200 ?M of each dNTP, 0.7 U DNA polymerase (Thermo Fisher Scientific, Hudson, NH, USA), 1 ? reaction buffer, 2 mM MgCl2 and 10 ng metagenomic DNA in a reaction volume of 20 ?L. Amplification was done with the PCR Thermal Cycler Multigene Optimax (Labnet, Edison, NJ, USA) using the following steps: (1) initial denaturation at 94 ¡C for 10 min; (2) 25 cycles of further denaturation at 94 ¡C for 45 s, annealing at 53 ¡C for 45 s, and extension at 72 ¡C for 1 min; (3) concluding extension at 72 ¡C for 10 min. The DNA from each sample was amplified three times, pooled, purified using the Fast Geneª Gel/PCR Extraction Kit (Nippon Genetics Europe GmbH, Japan), assessed for integrity by agarose (1%) gel electrophoresis and quantified with a Nanodropª 3300 Fluorospectrometer (Thermo Fisher Scientific Inc, Waltham, MA, USA.) using Quant-iTTM PicoGreenR dsDNA (Invitrogen, Carlsbad, USA). Equal amounts of the purified 16S rRNA gene amplicons were sequenced at Macrogen (Macrogen, Inc., Seoul, Korea), using paired-end (2?_?300 nt) Illumina MiSeq sequencing system (Illumina, USA). SRS2718854 SAMN08040967 T20.3 AMPLICON METAGENOMIC PCR Illumina MiSeq