Phloxine B Staining is Compatible With High-Throughput DNA Barcoding of Meiofauna

Area of Science:

• Marine Biology
• Molecular Ecology
• Biodiversity Science

Background:

• Integrative biodiversity research needs methods combining morphology and DNA sequencing.
• Meiofauna processing is bottlenecked by specimen sorting and stain interference with molecular workflows.

Purpose of the Study:

• To test phloxine B stain compatibility with DNA sequencing for meiofauna.
• To enable simultaneous morphological and molecular processing of meiofauna specimens.

Main Methods:

• Meiofauna were preserved in ethanol, propylene glycol, or DESS and stained with phloxine B.
• Specimens were manually isolated, and the 18S rDNA gene V1-V2 region was sequenced.
• Sequencing success was quantified across major meiofaunal phyla.

Main Results:

• Phloxine B staining did not inhibit DNA sequencing for nematodes, copepods, and annelids.
• High sequencing success was achieved regardless of preservative or staining duration.
• Platyhelminths showed lower success, potentially due to identification or primer issues, not dye inhibition.

Conclusions:

• Phloxine B is compatible with DNA amplification and sequencing, facilitating integrated data collection.
• This staining method enhances efficiency for processing microscopic taxa in biodiversity studies.
• The approach supports comprehensive, high-throughput biodiversity assessments and ecological monitoring.