EdU-Click on a Mock Community

Flow cytometry data on a mock community of bacterial isolates originating from the human body (<em>Escherichia coli </em>K12 (ATCC 25404), <em>Enterococcus faecalis</em> (SF24397), <em>Prevotella intermedia</em> (ATCC 25611), <em>Bacteroides fragilis </em>(32-6-I 11 MRS AN), and <em>Bifidobacterium longum </em>subsp. <em>infantis</em> (ATCC 15697), grown together or separately. The mock community was created by adding OD600 0.05 of the overnight cultures of each isolate to autoclaved reduced Bovine Heart Infusion broth (rBHI). The mock community was incubated for 1 hour, followed by another three hours with (or without) 20 uM 5-ethnyl-2'-deoxyuridine (EdU), a thymidine analogue, to capture the actively replicating cells. <br> The bacterial cells underwent a click reaction with Alexa Fluor 647 azide (APC channel) to make newly replicated bacteria fluoresece. They were also stained with Sybr Green I nucleic acid stain (FITC channel) to capture all bacteria. <br> In the file names: <br> BI =<em> Bifidobacterium longum </em>subsp<em>. infantis</em> <br> BF = <em>Bacteroides fragilis</em> <br> EC =<em> Escherichia coli </em>K12 <br> EF = <em>Enterococcus faecalis</em> <br> PI = <em>Prevotella intermedia</em>