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7 Comments
Emily Legan
10/27/2015 05:39:38 pm
All but one of my wells in the crystallization tray contained nothing. Condition #95 for Bacillus anthracis resulted in a precipitate of some sort. No, it was not a crystal, but I think there is potential. Perhaps we could see crystals if the tray is left alone for longer or the experiment is repeated with these specific conditions adjusted slightly.
Dr. Antony
10/28/2015 06:51:56 am
Emily,
Xinhang Yang
10/28/2015 08:33:56 pm
Maybe the time for SSB crystallization is not enough. Crystallization is a slow process. It's easy to get precipitate or microcrystal if we just try to reduce crystallization time like increase the concentration of SSB.
Dr. Antony
10/29/2015 06:34:00 am
Yes, we need to push the SSB concentration to ~ 100 uM for these trays. If not, we might be waiting for a long time.....FYI, we are currently between 30 - 45 uM in our experiments, except for one SSB prep where the concentration is ~120 uM. We will wait and see.
Rahul Sharma
11/6/2015 10:55:27 pm
After ~2 weeks of patience, the trays were finally analyzed for crystals. Unfortunately, I did not acquire any for Yersinia Pestis SSB. I found it interesting that some of my wells in the tray had large amounts of liquid present on the cover-slip, while others were completely dry. There didn't seem to be an inherent trend to this observation, but I wonder if my pipette precision had something to do with it. I also wonder why no supersaturation occurred. I think it has to do with the temperature of the environment, or possibly the amount of SSB protein added to the cover-slip. All in all the results were unsurprising, but a tad disappointing (especially the crystal false positive).
Nicholas Callard
12/4/2015 06:49:50 pm
I also worked with Yersinia pestis and found quite similar results. I really don’t think it was anything to do with your precision and instead more to do with concentrations that were used. I know that this experiment has the potential to be costly but it would have been super interesting to see if one of us was able to actually create a crystalized structure of the tetramer. Regardless before this my only knowledge of crystallization came from class discussions, so it was still very interesting to do the experiment. The dehydration component of the experiment never really made sense to me until I set up the experiment myself.
Tai Lu
12/7/2015 09:39:16 pm
I too worked had the Yersinia Pestis SSB but I did not see any crystallization after letting it sit there for a while. I wasn't sure because on some of the wells kind of looked like there were crystals present but after talking with the TA, Jessica, it was determined that it was only precipitation. If I had another chance of redoing this experiment, I actually would do it because I would like to find out if it was just an experimental error that I may have made or maybe Yersinia Pestis is just the way it is. Leave a Reply. |
AuthorDr. Edwin Antony and Dr. Sofia Origanti has put together this course. The material presented here are generated by the students of the BIOL 4102 class at Marquette University. (Fall 2015). Improvements can only be made if there is adequate feedback and we thank you in advance for your time. Archives
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