In this study, we implemented a modified metabarcoding approach to characterize the biome attached to polyethylene food bags after one month at sea in comparison to the free-living biome in two proximal marina and open water locations at the Mediterranean coast of Israel. To achieve higher taxonomic coverage and resolution we used five longer conventional barcode regions, targeting prokaryotes as well as a variety of different eukaryotic taxonomic groups. The barcodes were amplified and sequenced using the MinION nanopore platform with multiplexing and then subjected to an adapted bioinformatic pipeline to achieve optimal taxonomic characterization of the plastisphere compared to its environment.