Development of two new sets of PCR primers for eDNA metabarcoding of brittle stars (Echinodermata, Ophiuroidea)

Research Article

Metabarcoding and Metagenomics 7: e94298
https://doi.org/10.3897/mbmg.7.94298 (28 Feb 2023)

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Article title
Abstract
Key Words
Introduction
Materials and methods
- Design of new primer sets
- Water sampling from the tanks
- Water sampling from environmental water
- Library preparation and MiSeq sequencing with eDNA sample
- Decontamination in laboratory experiment
- Data preprocessing and taxonomic assignment
- Determination of target sequences of specimens collected from the study area
Results
- Development of 16SOph primers
- Threshold setting
Taxa detection from water eDNA
- Total reads
- Taxa detected with 16SOph1 from aquarium tanks and environmental waters around MMBS
- Taxa detected with 16SOph2 from aquarium tanks and environmental waters around MMBS
- Other detected taxa apart from Ophiuroidea
Discussion
- Usefulness of eDNA metabarcoding with 16SOph primers: 16SOph1 vs 16SOph2
- Taxa detected from eDNA metabarcoding – Aquarium tanks
- Taxa detected by eDNA metabarcoding – Moroiso
- Taxa detected by eDNA metabarcoding – Pier
- Taxa detected from eDNA metabarcoding – 80 m
- Taxa detected from eDNA metabarcoding – Deep sea (only 16SOph1)
- Future challenges – Optimization of eDNA metabarcoding
Conclusion
Data accessibility
Author contributions
Conflict of interest
Supporting agencies
Acknowledgements
References
Supplementary materials

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