2023 Early Hearing Detection & Intervention Conference

March 5-7, 2023 • Cincinnati, OH

5/22/2019 | 10:15 AM - 10:30 AM | HIGHLY ACCURATE FRESHWATER MACROZOOBENTHOS METABARCODING WITH NANOPORE MINION | 254 B

HIGHLY ACCURATE FRESHWATER MACROZOOBENTHOS METABARCODING WITH NANOPORE MINION

DNA barcoding coupled with High-Throughput Sequencing (HTS), commonly known as metabarcoding, has compelling advantage over traditional approaches for obtaining species distribution data. Although it has become a more commonly used tool complementing routine biomonitoring techniques, it is often difficult to detect all species diversity in a bulk sample. This can – in parts – be attributed to shorter read lengths most HTS instruments generate. Moreover, most HTS platforms are not portable, making in situ field-based sequencing infeasible. Oxford Nanopore sequencing platforms (ONT), such as MinION, are an exception. They are also known to provide longer reads albeit limited by rather high error rates (~12-15%). We used a freshwater mock community of 50 OTUs to test the capacity of ONT MinION coupled with rolling circle amplification (RCA) to provide long read metabarcoding results and estimated its diversity with >99% average sequence accuracy for 1D2 sequencing. Our DNA metabarcoding workflow of freshwater organisms using Nanopore sequencing shows that high error rates of long-read single-molecule sequencing can be mitigated using RCA. It appears that cost-effective field-based DNA metabarcoding is possible and future bioassessment programs will benefit from portable, accurate, species-level metabarcoding.

Bioassessment
Invertebrate
Monitoring

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Presenters/Authors

Bilgenur Baloglu (), Centre for Biodiversity Genomics (University of Guelph), bbaloglu@uoguelph.ca;