UltraClean 96 PCR Cleanup Kit (384)

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Publication protocol

Total DNA was extracted from frozen fecal pellets using the PowerSoil-htp 96 well DNA isolation Kit (MoBio) or the DNeasy PowerSoil HTP 96 Kit (QIAGEN). Barcoded primers were used to amplify the V3-V4 region of the 16S rRNA gene from extracted bacterial DNA using primers 515fB and 806rB via PCR (EMP). PCR clean-up was performed with UltraClean PCR Clean-Up Kit (MoBio) or UltraClean 96 PCR Cleanup Kit (QIAGEN) before quantification of amplicon products with Quant-iT dsDNA Assay Kit, high sensitivity (Invitrogen). Amplicons were pooled by adding 100 ng of each product. Illumina MiSeq paired-end reads runs were performed, with experiments divided among runs such that no more than 384 samples were read per run.


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Manufacturer protocol

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