16S Amplicons of Cladophora glomerata Microbiota: JD 185 Site Point Replicate 2
To remove planktonic microbial cells and unattached microfauna, as in previous studies of Cladophora microbiota (Zulkifly et al., 2012; Graham et al., 2015), algal filaments were washed three times in fresh autoclave-sterilized SD11, a defined medium closely resembling the chemical composition and pH of Lake Mendota water (Graham et al., 1996). All filaments of Cladophora were trimmed to the oldest 2 cm of basal cells near holdfasts, in order to consistently sample the colonized microbiota from equivalent locations, and avoiding newer, less highly colonized host cells at growing tips (as in Zulkifly et al., 2012 and Graham et al., 2015). The resulting algal materials were placed in 1.5 mL Eppendorf Safe-Lock tubes, and total genomic DNA was extracted on the same day using the DNEasy Plant Mini Kit (Qiagen) amended with 100킠 of 100mg/ml lysozyme (Sigma-Aldrich) solution to increase access to DNA of bacterial cells with recalcitrant cell walls (Yuan et al., 2012).