What question did this study set out to answer?

To investigate how tissue sampling methods, pooling strategies, PCR primer choices, and sequencing platforms influence species recovery in DNA metabarcoding of arthropod communities.

February 19, 2019Molecular Ecology ResourcesOpen Access

Metabarcoding a diverse arthropod mock community

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Authors

TBThomas BraukmannUniversity of Toronto NINatalya IvanovaNational Research University Higher School of Economics SPSean W. J. ProsserGenomics (United Kingdom)

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Implication

Experimental study reveals protocol and primer variations alter species detection in arthropod mock communities, highlighting the necessity of sufficient sequencing coverage.

Key Points

To investigate how tissue sampling methods, pooling strategies, PCR primer choices, and sequencing platforms influence species recovery in DNA metabarcoding of arthropod communities.
Assembled a mock community of 374 terrestrial arthropod species to test sequence recovery.
Evaluated four DNA extraction and pooling protocols: Bulk Abdomen, Bulk Leg, Composite Leg, and individually amplified Single Leg samples.
Sequenced amplicons across three high-throughput sequencing platforms: Illumina MiSeq, Ion Torrent PGM, and Ion Torrent S5, testing multiple primer sets.
Sequencing platform selection did not substantially alter overall species recovery, though Illumina MiSeq generated the highest sequence quality.
The Single Leg protocol achieved the most efficient species recovery due to low variation in amplicon abundance among taxa.
Among pooled-PCR treatments, Bulk Abdomen produced more uniform read abundance than Bulk Leg or Composite Leg, while primer choice significantly affected recovery and evenness.

Cite This Study

Braukmann et al. (2019) studied this question.

synapsesocial.com/papers/6a9efbd3cae4226ac4c36f97 https://doi.org/10.1111/1755-0998.13008

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Implication

Experimental study reveals protocol and primer variations alter species detection in arthropod mock communities, highlighting the necessity of sufficient sequencing coverage.

Metabarcoding a diverse arthropod mock community

Discussion

Member takes

Implication

Key Points

Cite This Study

Metabarcoding a diverse arthropod mock community

Discussion

Member takes

Implication

Key Points

Cite This Study

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