SRX1774741 MC1_D104 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446190 SAMN05007715 MC1_D104 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774742 MC1_D152 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446189 SAMN05007716 MC1_D152 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774743 MC1_D208 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446191 SAMN05007717 MC1_D208 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774744 MC1_D269 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446192 SAMN05007718 MC1_D269 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774745 MC1_D309 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446193 SAMN05007719 MC1_D309 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774746 MC1_D399 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446194 SAMN05007720 MC1_D399 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774747 MC1_D409 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446195 SAMN05007721 MC1_D409 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774748 MC1_D499 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446196 SAMN05007722 MC1_D499 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774749 MC1_D509 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446197 SAMN05007723 MC1_D509 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774750 MC1_D62 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446198 SAMN05007724 MC1_D62 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774751 MC2_D104 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446199 SAMN05007725 MC2_D104 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774752 MC2_D152 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446200 SAMN05007726 MC2_D152 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774753 MC2_D208 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446201 SAMN05007727 MC2_D208 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774754 MC2_D269 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446202 SAMN05007728 MC2_D269 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774755 MC2_D309 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446203 SAMN05007729 MC2_D309 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774756 MC2_D399 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446204 SAMN05007730 MC2_D399 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774757 MC2_D409 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446205 SAMN05007731 MC2_D409 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774758 MC2_D499 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446206 SAMN05007732 MC2_D499 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774759 MC2_D509 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446207 SAMN05007733 MC2_D509 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774760 MC2_D62 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446208 SAMN05007734 MC2_D62 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774761 MC3_D104 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446209 SAMN05007735 MC3_D104 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774762 MC3_D152 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446210 SAMN05007736 MC3_D152 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774763 MC3_D208 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446211 SAMN05007737 MC3_D208 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774764 MC3_D269 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446212 SAMN05007738 MC3_D269 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774765 MC3_D309 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446213 SAMN05007739 MC3_D309 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774766 MC3_D399 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446214 SAMN05007740 MC3_D399 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774767 MC3_D409 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446215 SAMN05007741 MC3_D409 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774768 MC3_D499 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446216 SAMN05007742 MC3_D499 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774769 MC3_D509 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446217 SAMN05007743 MC3_D509 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774770 MC3_D62 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446218 SAMN05007744 MC3_D62 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774771 MC4_D104 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446219 SAMN05007745 MC4_D104 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774772 MC4_D152 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446220 SAMN05007746 MC4_D152 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774773 MC4_D208 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446221 SAMN05007747 MC4_D208 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774774 MC4_D269 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446223 SAMN05007748 MC4_D269 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774775 MC4_D309 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446222 SAMN05007749 MC4_D309 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774776 MC4_D399 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446224 SAMN05007750 MC4_D399 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774777 MC4_D409 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446225 SAMN05007751 MC4_D409 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774778 MC4_D499 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446226 SAMN05007752 MC4_D499 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774779 MC4_D509 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446227 SAMN05007753 MC4_D509 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium SRX1774780 MC4_D62 SRP075403 PRJNA322031 "Construction protocol: The v6-v4 hypervariable regions of the bacterial SSU rRNA gene were amplified using degenerate primers. The primers were 1064R (CGACRRCCATGCANCACCT) and 518F (CCAGCAGCYGCGGTAAN) fused to Roche GSFLX amplicon sequencing adapters and including 5 nt multiplexing barcodes. We generated PCR amplicons in triplicate 33 uL reaction volumes with an amplification cocktail containing 1.0 U Platinum Taq Hi-Fidelity Polymerase (Invitrogen, Carlsbad, CA), 1X Hi-Fidelity buffer, 200 uM dNTP PurePeak DNA polymerase mix (Pierce Nucleic Acid Technologies, Milwaukee, WI), 1.5 mM MgSO4 and 0.2 uM of each primer. We added approximately 10-25 ng template DNA to each PCR and ran a no-template control for each primer pair. Amplification conditions were: initial 94C, 3 minute denaturation step; 30 cycles of 94C for 30s, 60C for 60s, and 72C for 90s; final 10 minute extension at 72C. The triplicate PCR reactions were pooled after amplification and purified using Ampure beads. Purified DNA was eluted in 30 uL of Qiagen buffer EB. We assessed the quality, size and concentration of PCR products on a Perkin Elmer Caliper GX. Reads were demultiplexed and barcodes removed for submission." SRS1446228 SAMN05007754 MC4_D62 AMPLICON METAGENOMIC PCR 535 0 Application Read Forward 1 454 GS FLX Titanium