Experimental quantification of pollen with DNA metabarcoding using ITS1 and trnL.

摘要: Although the use of metabarcoding to identify taxa in DNA mixtures is widely approved, its reliability quantifying taxon abundance still subject debate. In this study we investigated relationships between amount pollen grains mock solutions and high-throughput sequence reads how relationship was affected by counting methodology, number PCR cycles, type markers plant species whose have different characteristics. We found a significant positive sequences solutions. However, better were obtained with light microscopy as grain method compared flow cytometry, chloroplastic trnL marker ribosomal ITS1 30 when 25 or 35 cycles. provide list recommendations improve quantification.