Metabarcoding Presentation By: Anna, Sharleen, Maureen

Type of Article Review article No methods or results sections Broad overview of what was found when the sequencing analyzed Acknowledgment section Lots of sources being cited throughout the article

Purpose Realized the large diversity of soil parasitic protists Many ways to help find this diversity Used to just have people with taxonomic expertise now more advanced technology HTS (high-throughput sequencing) There continues to be improvements on the sequencing technology - Finding this diversity helps ecological functions and the ecosystem

Metabarcoding Challenges The majority of the few studies investigating the community structure of soil protists applied time-consuming microscopic determinations that require profound taxonomic expertise. Another challenge is finding a diversity of soil protists similar to that of bacteria and having their sequence closely match it to that of parasites There are little studies known about the community structure of soil protists so the studies made on soil protists require profound taxonomic expertise

Information that can be applied to the Research Question (How to extract DNA and what primer we should use to amplify it?) Problems: - “Close attachment of many protists to soil particles eliminates straight forward extraction.” -“Designing universal protist-specific primers is impossible”. -The diversity of the protists is just too great and too different -

Eukaryotic Tree of Life

Information that can be applied to the Research Question (How to extract DNA and what primer we should use to amplify it?) Solutions: -The trick is using “multi-methodological approaches” to link -morphological -molecular -functional information of distinct parasites,

Information that can be applied to the Research Question So we can: -decipher patterns -determine identities -evaluate the effects of the treatments on the different protists. (How to extract DNA and what primer we should use to amplify it?)

Information that can be applied to the Research Question (How to extract DNA and what primer we should use to amplify it?) We can use: - Classical cultivation based efforts - Manually mined DNA (Amplicon) - based HTS studies - Phylogenetic analyses - PCR free Metatranscriptomic and Metagenomic approaches *Most reliable for soil protist communities* - pH Analyses

Our Questions Vocabulary: Concepts: Phylogenetic-“relating to the evolutionary development and diversification of a species or group of organisms, or of a particular feature of an organism.” Amplicon-“a piece of DNA or RNA that is a source and/or product of natural or artificial amplification or replication events.” Monophyletic- “(of a group of organisms) descended from a common evolutionary ancestor or ancestral group, especially one not shared with any other group.” Concepts: HTS ( the detailed process and the meaning of the results found)

Bibliography Ph Picture: https://www.google.com/search?q=ph+test&rlz=1C1NDCM_enUS720US720&source=lnms&tbm=isch&sa=X&ved=0ahUKEwjjt_vL2uDYAhUPLqwKHcn_AWEQ_AUICygC&biw=1280&bih=540&dpr=3#imgrc=vdTd5Ka0XymYrM: Stentor and Tetrahymena gifs: https://www.google.com/search?q=stentor+gif&rlz=1C1NDCM_enUS720US720&source=lnms&tbm=isch&sa=X&ved =0ahUKEwjIsaiz2-DYAhVQA6wKHUBxDV4Q_AUICigB&biw=1280&bih=540#imgrc=3Cf3MwzF3AL60M: https://www.google.com/search?rlz=1C1NDCM_enUS720US720&biw=1280&bih=540&tbm=isch&sa=1&ei=wCdgW sHOEIPysQXYgq_oBQ&q=tetrahyemnea+gif&oq=tetrahyemnea+gif&gs_l=psy- ab.3..0i13k1.53481.57768.0.57924.16.16.0.0.0.0.69.853.16.16.0....0...1c.1.64.psy- ab..0.16.851...0j0i67k1.0.ptkt0kWG8yQ#imgrc=wXG1ntbmfJvfkM: Ciliate Morphology and Classification Pictures: https://www.slideshare.net/eukref/zhang-eukref2015

Bibliography Title Gif: https://www.google.com/search?rlz=1C1NDCM_enUS720US720&biw=1280&bih=589&tbm=isch&sa=1&ei= rbZfWqOSBcyWtQWroK8g&q=soil+ciliates+gif&oq=soil+ciliates+gif&gs_l=psy- ab.3...207466.207871.0.208032.4.4.0.0.0.0.50.164.4.4.0....0...1c.1.64.psy- ab..0.0.0....0.XUSBX_c4yt4#imgrc=Df3KVsTQDjIMSM: