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_Isolation/Culturing Efforts_:  The data from Aims 1 and 2 will be used as a benchmark to determine how culturing efforts are going and to refine efforts to get specific microeukaryotes of interest.  A variety of media recipes found previously successful at isolating seagrass-associated fungi will be used here. These include potato dextrose agar, malt-dextrose agar, glucose peptone yeast agar and cornmeal agar, with all media being supplemented with salt and antibiotics (27, 28, 42). Media will also be made using Z. marina as the carbon source as was done by Torta el al (27) using the seagrass, Posidonia oceanica.   To enrich for amoeba, samples will be inoculated onto plates containing ???? media (1 mL 0.5 M MOPS buffer, 0.1 g yeast extract, 0.1 g tryptone, 15 g agar and 1X seawater base to make 1L) with a marine bacterium as a food source. Upon formation, plaques will be picked from the plate and inoculated into flasks containing approximately 10 mL of liquid media (no agar added). Flasks will be monitored for amoeba growth. 
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-modified hay infusions with seagrass for HS outreach  _Molecular Biology_:  The V4 and V9 regions of the 18S rRNA gene will be amplified as in Aim 1 to identify isolated organisms and Sanger sequenced. Sequences with homology will be found using the SILVA database and Genbank searches. Phylogenies will then be built using RAxML and microeukaryotic taxonomies will be assigned based on tree position.  Multi species: same methods as Zmarina