The abundance, biomass, and composition of phototrophic picoeukaryotes (PPE, cell size less than 3 μm) were studied in Onega Bay of the White Sea in June 2015. The highest PPE abundance and biomass were registered in the 0–5-m water layer. In the bay, in the 0–5-m water layer, the average abundance and biomass varied from 0 to 36.8 × 104 cell/L and from 0 to 117 μg С/m3, respectively. The Illumina sequencing of V4 region of 18S rRNA gene revealed eight classes of PPE. Mamiellophyceae dominated both by number of reads and by operational taxonomic units. The green algae Bathycoccus prasinos, Ostreococcus tauri, and Micromonas pusila, as well as diatoms Skeletonema marinoi and Minidiscus trioculatus, were identified to the species level.