experiments for reassurance
- Posted by darkeye on May 28th, 2008 filed in experiments, log
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As the last few experiments didn’t really work out, it’s time to replicate some of the originals, and re-think the ones did recently. Starting from square 1:
- re-grow the bacteria on top of a plate, taken from the cryogenic storage
- create overnight liquid cultures from single colonies of the regrown bacteria
- create agar plates with the liquid culture in them
- perform arabinose experiments with it
Moreover, the UV-light bacteria killing experiment had to be re-done, as most probably the plastic cover of the petri dish itself filters out the harmful UV lights 🙂 So I just did that, and have some plates exposed to UV lights without the petri dish cover, and now waiting in liquid LB to see if they grow out.
In more detail:
day 1: re-grow bacteria
Re-grow the bacteria on top of a plate, taken from the cryogenic storage. I did this on two plates today, one with just ampicillin in it, and the other also with arabinose, to see if it turns fluorescent at all. Easy reassurances create a flow experience 🙂
day 2: create liquid cultures
Take single colonies from the plates that grew overnight, and create liquid cultures from them. Also verify that the plate with the arabinose in it turned fluorescent.
day 3: create agar plates & perform arabinose experiments
Take the liquid culture that grew out and crate agar plates with the bacteria in them. As the plates are a few (at least 3) hours old, drop 5μl droplets of arabinose on them, and leave overnight to develop. Do the experiment in the following three settings:
- always have the plate in a 37°C incubator
- always leave the plate at room temperature
- always put the plate in the fridge
Moreover, have two kinds of plates for each experiment:
- regular LB-agar plates with bacteria in them
- ‘top agar’ plates
day 4: verify fluorescent pixels
Take the plates that had the arabinose dropped on them, and verify if any of the drops turned fluorescent, or the whole plate turned fluorescent.
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