anniedoyle.bsky.social
Microbiologist studying C. difficile toxins | PhD candidate @BallardLab | NIAID NRSA F31 Fellow
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Right there with you
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We have an eppendorf thermomixer that is awesome. You can swap the top to do reg eppendorfs, 96 well, 384 well, etc. Different models come with different adapters.
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I feel like that’s true for a lot of gut microbes that health focused labs grow. Most people don’t even know how to do roll tubes these days so that’s pretty depressing.
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😂 listen, I took a few steps back from looking at the host in order to investigate some groovy bug stuff. I’ll find my way back to the fancy scopes one day!
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To be 100% honest, I really don’t consider C. diff to be a true anaerobe but rather an aerotolerant bacteria since it doesn’t immediately die when oxygen is a mile away like most strict anaerobes I’ve worked with… maybe it’s just extra good at making superoxide dismutase!
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But we don’t gas exchange with a manifold like I have with acetogens, so there must be a microaerophilic environment at the very top of the dialysis tubing, most likely stimulating the superoxide dismutase
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I have a strong feeling that’s exactly what that is. For growing native toxin we actually don’t grow it in the chamber but instead boil, purge, and stopper 4L flask then inject culture into large amounts of hanging dialysis tubing and harvest after a few days
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Nope! I feel like maybe this could be a copper containing protein maybe? I’m keeping the column in the cold room for now because I want to go back and try to figure this out.
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I guess it would depend on the stability and flexibility of your protein…but I would go for CryoEM first and just up the thickness of the ice a little bit if you are struggling to trap different orientations
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This is truly a work of art
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🤞
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Looks like potential mini preps checking colonies for whether or not the transformations are good?
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Oh no!! I hope you have a speedy recovering!