Evaluation of full-length 16S rRNA amplicon sequencing using Oxford Nanopore Technologies for diversity surveys of understudied microbiomes

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Abstract

  • The use of long-read sequencing using portable Oxford Nanopore Technologies (ONT) is becoming increasingly popular in the study of host-associated microbiomes. However, its application has not yet been optimized for characterizing understudied microbiomes, such as those in marine environments.

  • We evaluated the accuracy and consistency of ONT sequencing of full-length 16S rRNA genes for diversity surveys of symbiotic dinoflagellate (family Symbiodiniaceae) microbiomes.

  • When comparing amplicon sequencing of the full-length bacterial 16S rRNA gene with only its V4 hypervariable gene region using a known microbial community, the former recapitulated the bacterial taxonomic composition more accurately.

  • ONT sequencing was also highly consistent between sequencing runs and flow cells.

  • Long-read sequencing technologies enable microbiome surveys using the full-length 16S rRNA gene, achieving higher accuracy and resolution.

  • This work validates ONT long-read sequencing as a powerful tool for marine microbiome studies to catalyze advancements in the fields of ecology and evolution, resource management, and conservation.

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