Summer Reflections

In reflection of all that has happened this summer, I’ve realized that the past 10 weeks have gone by incredibly quickly.  Overall, I was able to test samples from four different gels and find a plethora of peptides and subsequent proteins.  In addition, I learned a lot about the other projects we are doing in the lab, like our kinetic method projects for finding proton affinity of unnatural amino acids.  We also flowing afterglow mass spectrometer up and running and taking data, something that has not been attained in our lab for several years.  There was a lot of progress in both my project and the others in our lab, despite the power outage and lost weeks.

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The Flowing Afterglow

Aside from my main project, I have been helping get another mass spectrometer in the lab up and running: the flowing afterglow.  This instrument allows us to conduct reactions in the flow tube to produce ions using volatile compounds.  In addition, temperature and pressure can be controlled in the instrument to create an atmospheric environment for these reactions.

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Learning about SEQUEST

The program that we use in order to identify proteins is called SEQUEST.  When I first started working in the lab, I ran spectra through this program by changing various parameters and then watching it work, without really understanding the process that it takes to come up with the best peptide/protein matches for the sample.  Now I understand the modifications of the program help get the most accurate values possible.

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Long time, no see!

It’s been quite a while since I’ve been able to update my progress due to some unforeseen issues.  On the 19th of June, William and Mary mistakenly cut the power to the Integrated Science Center as a part of a scheduled campus-wide power outage.  When the power was reintroduced to the buildings, the instruments, which previously were on, turned back on immediately and improperly.  This blew both power supplies to the mass spectrometer, which can only be replaced by an outside repairperson.  During the three week period of waiting for him to find replacement parts and travel from North Carolina to Williamsburg, I did a lot of research for our kinetic method projects, namely condensing information about experimentally and theoretically retrieved proton affinity values for glycine.  In addition, we worked on our flowing afterglow mass spectrometer, which needed some plumbing adjustments and a change to the software before being able to produce scans.

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Splitting to Optimize Flow Rate

For the past several weeks, the lab has made progress in prepping for the summer and fall. For our kinetic method experiments, we were able to sort, clean, and inventory all samples for easy access including general inventory around the lab.  The pump for the flowing afterglow mass analyzer in the lab has finally been reinstalled (in it’s 800 pounds of glory), pump oil has been changed for all pumps, and gas tanks have been replaced for all of the instruments.  Now in week four, I have been able to start looking at the instrument my project uses and how to improve it so we can start running protein samples.

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