eDNA Metabarcoding
To test the utility of the FishCARD database, we metabarcoded fish eDNA
from 3 sites off Eastern Santa Cruz Island, CA in 2017. We collected
seawater samples from 10m depth using a 4 L Niskin bottle. From this
sample, we gravity filtered 1 L through a 0.2 µm Sterivex (Millipore
Sigma, Burlington, MA, USA) filter in the field and preserved on ice
before being transported to a -20˚C freezer. We then extracted eDNA from
the filters using the modified Qiagen DNeasy Blood and Tissue Kit
extraction protocol of Spens et al. (2017), and then amplified it via
PCR using the MiFish Teleost primers and thermocycler profile above. We
prepared libraries following the methods of Curd et al. (2019) and
sequenced these samples on an Illumina MiSeq PE 2x300 at UCLA Technology
Center for Genomics and Bioinformatics (see Supplemental Methods for
detailed library preparation protocol).