Polymerase and primer biases in eukaryote ultra-long-read metabarcoding

In PacBio ultra-long-read metabarcoding of eukaryotic mock communities and 10 soil samples, Solis or Fusion polymerases and Euk575F-21Rngs primers best balanced data quality and biodiversity recovery.

• Why it matters: Ultra-long-read metabarcoding depends on high-processivity polymerases and primer pairs that span most of 18S, ITS, and 28S, but their relative suitability and bias are not well characterized.
• What they did: The study compared five commercial high-processivity polymerase kits and three ultra-long primer pairs against a full-ITS primer pair, using two mock communities and 10 soil samples with PacBio sequencing.
• The result: Polymerase and primer choices produced large differences in artefacts and biodiversity; for soil eukaryotes, Solis or Fusion with Euk575F-21Rngs is recommended, and ultra-long amplicons recovered substantially lower biodiversity than medium-sized full-ITS amplicons.

AI summary of the abstract · may contain errors · not medical advice · read the paper.