Evaluation of Filtration and DNA Extraction Methods for Environmental DNA Biodiversity Assessments across Multiple Trophic Levels.

摘要: Metabarcoding of marine environmental DNA (eDNA), the suspended in seawater originating from tissue, cells, or extracellular DNA, offers opportunity to survey biological composition communities across multiple trophic levels a non-invasive sample. Here we compare results eDNA metabarcoding same samples collected kelp forest Monterey Bay, California order establish methods for future cross-trophic level analysis. Triplicate 1 L water were filtered using five different 47 mm diameter membrane filters (PVDF, PES, GFF, PCTE, and NC) was extracted triplicates each filter-type three widely-used extraction (the DNeasy Blood Tissue kit, MoBio PowerWater Isolation standard phenol/chloroform methods) resulting 45 individual prepared with 15 workflow combinations. Each extract amplified PCR primers 16S rRNA gene (microorganisms; Bacteria Archaea), 18S (phytoplankton), 12S (vertebrates) genes, products sequenced on an Illumina MiSeq platform. The richness community microbial, phytoplankton, vertebrate OTUs not significantly between any 0.2 μm pore-size filter types kits. However, resulted structures. This study provides insight into choices filtration use biodiversity assessment single We recommend combination either PVDF, NC cross comparison.