Optimizing Methods for Insect Metabarcoding
Due to its efficiency, metabarcoding is widely used for molecular identification of organisms. We sampled insect diversity using light traps in lowland dipterocarp forest of Tengku Hassanal Wildlife Reserve in Pahang, Malaysia. Each light trap sample was homogenized and repeatedly subsampled to identify the number of subsamples required to detect total estimated insect OTU diversity in a light trap. Insect OTU diversity from 72 subsamples (12 subsamples from each of six light traps) was characterized by sequencing part of the mitochondrial Cytochrome Oxidase I gene. We found four and eight 100 uL subsamples sufficient to detect at least 90% and 95%, respectively of insect OTU diversity collected in each light trap regardless of a range of variation in wet weight (137.9-445.4 g).