Assessing the lysis of diverse pollen from bulk environmental samples for DNA metabarcoding

Pollen is ubiquitous year-round in bulk environmental samples and can provide useful information on previous current plant communities. Characterization of pollen has traditionally been completed based morphology, requiring significant time expertise. DNA metabarcoding a promising approach for characterizing from samples, but accuracy hinges successful lysis grains to free template DNA. In this study, we assessed the morphologically taxonomically diverse one most common sample types metabarcoding, surface soil. To achieve this, four species artificial mixture was spiked into soils collected Colorado, North Carolina, Pennsylvania, subsequently subjected extraction using both PowerSoil Pro Kits (Qiagen) with heated incubation (either 65 °C or 90 °C). Amplification Illumina sequencing internal transcribed spacer subunit 2 ( ITS2 ) duplicate each (total n, 76), resulting reads identified GenBank. The Kit statistically outperformed total yield. When either kit, temperature (65 °C) used had no impact recovery DNA, amplicon sequence variants (ASVs), reads. This study highlighted that feasible commercially available kits, downstream be accurately characterize such types.