Isolation and barcoding of pollen DNA and trace pollen-free DNA using next-generation sequencing for honey authentication

We have explored the DNA-based characterization of honey origins using next-generation amplicon sequencing targeting the nuclear ribosomal ITS2 region of plant genomic DNA. We isolated pollen DNA and trace pollen-free DNA from five honey samples originating from five countries. The trace pollen-free DNA from filtered honey samples was isolated using three types of adsorbents: (i) anti-dsDNA antibodies coupled to magnetic microspheres; (ii) Q Sepharose anion exchange adsorbent; and (iii) ceramic hydroxyapatite, CHT Type I. Among the three methods tested, we observed the highest similarity between DNA sequences obtained from NGS of pollen DNA and pollen-free DNA isolated using the Q Sepharose method.