Metabarcoding or high-throughput sequencing of a specific genetic marker is a powerful technique, widely used today, to analyze biodiversity across distinct environments and taxonomic groups. Plankton ecologists have benefited tremendously from the growing accumulation of metabarcoding studies. Novel biogeographic patterns have been established by the analysis of datasets from the Tara Oceans and Ocean Sampling Day projects. Novel lineages without cultured representatives have been uncovered. This chapter begins by going back to the steps that led Carl Woese and George Fox to define the concept of “molecular marker.” Among the multitude of exciting findings brought by high-throughput sequencing technologies, perhaps the major impacts are found in the study of picocyanobacteria and microbial eukaryotes from plankton communities. We then detail the different steps and choices that are involved in designing, performing, and analyzing a metabarcoding study. We are using a compilation of about 250 metabarcoding studies to present the major trends in terms of the gene marker used and environment probed. An alternative approach to metabarcoding developed for marine picocyanobacteria is also briefly discussed. We are then focusing on specific habitats and processes that have benefited from metabarcoding: the study of polar ecosystems, the functioning of the marine biological carbon pump, predator-prey interactions, and picoeukaryotic phytoplankton in highly urbanized lakes. Finally, we offer some perspectives on emerging trends, such as the use of metabarcodes combined with supervised machine learning for biomonitoring, the link between metabarcoding and functional diversity in trait-based studies and the massive sequencing of long DNA fragments.