RT Journal Article T1 Evaluation of DNA extraction methods and bioinformatic pipelines for marine nano- and pico-eukaryotic plankton analysis A1 Muñoz Colmenero, Ana Marta A1 Sánchez, Ana A1 Correa, Begoña A1 Figueiras, Francisco G. A1 Garrido, José Luis A1 Sotelo, Carmen G. AB The smallest size fractions of plankton, nano- and pico-plankton, have been highlighted due to they accomplish key functions in marine ecosystems. However, the knowledge about some of them is scarce because they are difficult or impossible to be detected and identified with non-DNA-based methodologies. Here we have evaluated five DNA extraction protocols (MT1–MT5) and seven bioinformatic pipelines (P1–P7) to find the best protocol for detecting most of the eukaryotic species of nano- and pico-plankton present in an environmental sample using Ion Torrent technology. The protocol MT3 was the most reproducible methodology, showing less variation among samples, good DNA quality and sufficient quantity to amplify and sequence the eukaryote species, offering the best results after sequencing. For bioinformatic analyses, P1 and P7 resulted in the highest percentage of detection for the difficult-to-detect species in mock communities. However, only P1 avoided the confusion with other closed species during the taxonomic assignment. The final protocols, MT3-P1 (free) and MT3-P7 (private), showed good and consistent results when they were applied to an environmental sample, being a valuable tool to study the eukaryotes present in environmental samples of nano- and pico-plankton, even for the genera that are difficult to be detected by other techniques. PB Frontiers Media SN 2296-7745 YR 2021 FD 2021-01 LK https://hdl.handle.net/20.500.14352/109658 UL https://hdl.handle.net/20.500.14352/109658 LA eng NO Muñoz-Colmenero M, Sánchez A, Correa B, Figueiras FG, Garrido JL, Sotelo CG. Evaluation of DNA Extraction Methods and Bioinformatic Pipelines for Marine Nano- and Pico-Eukaryotic Plankton Analysis. Front Mar Sci 2021;7:584253. https://doi.org/10.3389/fmars.2020.584253. NO MM-C holds a “Juan de la Cierva Formación” postdoctoral fellowship from the Spanish Ministry of Education with reference code FJCI-2017-32722. NO Ministerio de Ciencia e Innovación (España) NO Ministerio de Educación (España) DS Docta Complutense RD 23 abr 2025