Universal Metabarcodes (16S & 18S) from the Australian Antarctic Division RV Aurora Australis 2015/16 Voyage 3 - K-AXIS - (Kerguelen Axis)

This metabarcoding dataset is derived from PCR amplification of marine plankton-derived DNA from the Kerguelen-Axis Marine Science program in 2016. Metabarcoding reads were generated using a single primer set that simultaneously amplifies both 16S and 18S genes, though users should note that specific bioinformatic procedures are required to recover and analyze 18S sequences (see: https://github.com/jcmcnch/eASV-pipeline-for-515Y-926R).This dataset is part of a larger collaborative project called GRUMP (Global rRNA Universal Metabarcoding of Plankton) which has produced metabarcoding data from worldwide cruises from the same universal primer set (Parada et al., 2016, doi:10.1111/1462-2920.13023) on unfractionated samples (> 0.2 um). This primer set perfectly matches the rRNA of most surface ocean organisms, including eukaryotic and metazoan 18S (McNichol et al., 2021, doi:10.1128/msystems. 00565-21). As a result, the sequences here represent a full-community profile of each water sample with the same denominator and the same primer set (Yeh et al., 2021, doi:10.1111/1462-2920.15553).Notes for data users:Only limited environmental covariate data has been uploaded with the raw sequences. More data will be made available at the locations specified below.Final, processed data (16S + 18S relative abundances with taxonomic annotations) will be provided through the Simons Foundation CMAP (Collaborative Marine Atlas Project; https://simonscmap.com/) alongside environmental covariates. If you do not wish to reanalyze these data, we suggest using this data product.Bioinformatic intermediates, and scripts are stored at OSF in a single umbrella repository (https://osf.io/57dpa/). This is a useful place for those who might wish to analyze only a subset of our data (e.g. 18S or 16S only) or who wish to understand the bioinformatic processing in greater detail.Additional updates (e.g. linking additional environmental covariates to metabarcoding data) will be provided at our github page (https://github.com/jcmcnch/Global-rRNA-Univeral-Metabarcoding-of-Plankton). This is a good place to check for the latest updates to the GRUMP project.Specific notes for this dataset:This dataset contains relative abundance and taxonomic information for planktonic organisms collected by filtering whole seawater (2L volumes) collected from between 5 and 4625 m depths onto 0.2 um Sterivex polyethersulfone membrane filters (Millipore, cat# SVGPL10RC) during the Kerguelen-Axis Marine Science program in 2016. Samples were collected at sea by Bruce Deagle and Lawrence Clarke (Australian Antarctic Division) as part of the Australian Antarctic Division RV Aurora Australis 2015/16 Voyage 3. Samples were stored after excess liquid was purged at -80C until analysis. DNA was isolated and purified by Swan LS Sow and Bronwyn Holmes in 2017 (Environmental Genomics Team, CSIRO Environment) using a modified phenol:chloroform:isoamyl based extraction protocol of the DNeasy PowerWater Sterivex Kit (MoBio-Qiagen, Hilden, Germany) (Appleyard et al., 2013; Brown et al., 2018, Sow et.al., 2022). DNA was amplified, and sequenced by Jesse McNichol, Yubin Raut, & Bruce Yanpui Chan (Fuhrman Lab, USC) in 2020 and 2021.PCR amplification and sequencing:5' master mix (product 2200400/2200410) was used for DNA amplification with the 515Y (5'-GTGYCAGCMGCCGCGGTAA) and 926R (5'-CCGYCAATTYMTTTRAGTTT) primers with Illumina adapters and barcodes pre-ligated (as noted here: dx.doi.org/10.17504/protocols.io.vb7e2rn). Sequencing was done at Tufts University Medical School using HiSeq Rapid Run technology (2x250 bp).