Box 1: Designing a study using pollen DNA metabarcoding and related methods
There is a broad range of applications for pollen DNA metabarcoding and related methods. The experimental design will depend on the questions being addressed in the study and the time, budget, and skillsets available. There are four main steps in identifying plant species from pollen: (1) obtaining the pollen sample; (2) selecting appropriate gene region(s) and creating a reference database; (3) isolating and sequencing DNA; (4) bioinformatic analysis to compare sequencing reads to reference databases (Fig. 2). These four steps are outlined below.