16s rRNA of positive control
Earth Microbiome Project protocols and primers for amplification of both the 16s V4 region and library prep. PCR product size was determined by visualization with the TapeStation HS DNA tape and quantified using the protocol outlined in the Illumina Library qPCR Quantification Guide. PCR product quantity was double checked using the Qubit fluorometer, and samples that failed to show quantities more than the blank used in the standard curve (0.305ng/uL) were excluded from sequencing steps.. Libraries were normalized to the lowest concentration >1ng/uL, then pooled. Any samples <1ng/uL were added to the pool without normalization. The libraries were hen sequenced at the CQLS on Illumina MiSeq (300bp paired-end).