Endophyte isolation followup

 

A few samples were selected for hyphal tipping and pure culture isolation.  Shown, in no particular order, are some of the samples to be taken and re-plated on fresh PDA.   Small pieces of the mycelia were excised with a sterile scalpel and placed in the center of fresh plates.  In some instances, like the dark red pigmented colony growing within a few others, hope of a perfectly pure inoculum wasn’t expected, and instead we were just interested to see what might grow on the plate.

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There were instances like the first picture, in which a very homogeneous mycelial mass was obtained, which looks like a pure culture isolation.  We will perform DNA isolation and ITS amplification on some of the select organisms, as well as observe some of them under the microscope.  This week, agar plugs of pure strains will be put into liquid broth to ferment, with the downstream goal of processing the fermentation broth to isolate a crude extract of secondary metabolites to be used for bio-assay guided fractionation.

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