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.