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URL: https://dx.doi.org/10.17504/protocols.io.mv2c68e

Authors: Collin Closek, Anni Djurhuus, Katie Pitz, Ryan Kelly, Reiko Michisaki, Kristine Walz, Hilary Starks, Francisco Chavez, Alexandria Boehm, Mya Breitbart

Group: MBON eDNA

Summary: This protocol is aimed at amplifying the 18S rRNA hypervariable region 9 (18S V9) in eukaryotes with a focus on microbial eukaryotes. Amplicons generated using this protocol can then be sequenced using the Illumina platform. The primers (1391F, EukBr) utilized in this protocol are based on the primer utilized in Amaral-Zettler et al 2009 and the Earth Microbiome Project (EMP).Citations Amaral-Zettler LA, McCliment EA, Ducklow HW, Huse SM (2009) A Method for Studying Protistan Diversity Using Massively Parallel Sequencing of V9 Hypervariable Regions of Small-Subunit Ribosomal RNA Genes. PLoS ONE 4(7): e6372. doi:10.1371/journal.pone.0006372 Earth Microbiome Project - http://www.earthmicrobiome.org/emp-standard-protocols/18s/

Affiliations: Center for Ocean Solutions, Stanford University, CA, University of South Florida, College of Marine Science, St Petersburg, FL, Monterey Bay Aquarium Research Institute, Moss Landing, CA, University of Washington, Seattle, WA, Monterey Bay Aquarium Research Institute, Moss Landing, CA, Monterey Bay Aquarium Research Institute, Moss Landing, CA, Center for Ocean Solutions, Stanford University, CA, Monterey Bay Aquarium Research Institute, Moss Landing, CA, Center for Ocean Solutions, Stanford University, CA, University of South Florida, College of Marine Science, St Petersburg, FL

Version: 1

Publication Date: 2018

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