Hi fellow protein crystallographers,
I'm wondering if some of you kind souls could take a look at some peculiar sitting droplets that I've acquired from a crystal screen to help me understand what I'm seeing!
I'm fairly inexperienced with growing crystals, but I'm attempting to crystallise a protein (~70kDa) for the first time, I've set up a few screens using commercial products and have a variety of results (some very promising!).
I was really interested to see the following effects in quite a few of my drops (~10 out of 576), effectively showing spindles extending from the droplet out into the rest of the well, along with some dark spotting. Initially I thought that maybe some bacteria might be growing in the droplet, but the spindles that extend out of the drop are in a few different conditions.
Does anybody know what this effect is, and what the dark spots are? They don't look very similar to nucleation points in my other conditions.
As an aside, if anybody has a practical guide for protein crystal growing that they could recommend, that would be wonderful!
Thanks a lot,
Rob
That looks like fungal growth in your drops. To prevent this, you may want to filter all your 'sensitive' solutions used for crystallization setups, at least your buffers.
Hi Tjaard Pijning.
Oh dear, that's a bit concerning! Our buffers were taken from commercial screen solutions that come pre-filtered in a deep 96 well plate, so it could be that some of our screen solutions have contaminants. Perhaps it's time to replace them.
Thanks a lot,
Rob
Hi Rob,
I agree with Tjaard Pijning. As a structural biology researcher, I can tell you directly that the reservoir was contaminated. If you used commercial buffers, it is recommended that you replace them and best to aliquot before use. And if you used buffers prepared by yourself, please filter before use.
All the best!
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