7/16/07

Today I divided my time between Fromme's lab and Wachter's lab. In Wachter's lab, we are isolating GFP through the use of primers and a broth in E. coli with an antibiotic that the plasmid is resistant to. In Fromme's lab, we were trying to replicate experiments creating inorganic crystals using the materials that would be typical of a high school lab and Dr. Fromme commented on how much more difficult simple experiments then become. She showed us a particular catalog that would allow us to purchase better equipment- not lab quality- but certainly above rudimentary for our work creating copper sulfate crystals. We found out how to apply the terms saturated and supersaturated in the lab setting as well and will be meeting once again with Dr. Fromme at 2:30 PM to review our powerpoint thus far.

7/13/07

Today was rather stressful and we were trying to figure out our presentation. We listened to a grad student present his current data very succinctly- since there were no questions and he was a physics person amidst chemists. We also ate more cake and then tried to figure out what would be best to do for next week in light of our powerpoint and poster session. As such, there was a bit of disagreement between our research groups so we decided to split up. Fortunately, Petra Fromme came to see us and talk with us about how best to do our powerpoint so we had a much better idea and it will be started over the weekend as we have divided the tasks.

7/12/07

We finished working in the cold room to perform macroseeding for PSI crystals which required us to have the hands and steadiness and concentration of a surgeon. I loved b eing in a "different" environment at 4 degrees Celsius and I found it unusual and exciting. Additionally, I got more time to speak with Petra even more on a personal level and find out about some of her amazing "science" stories. We then took her out to lunch and continued to talk about the possibility of having some of our students working with her- networking at its best. Additionally, I believe the professors are very impressed and grateful for the work we have done and the feeling is very much mutual.

Pics

Our DNA! GFP!

7/11/07

Today, we continued work on PCR and gel electrophoresis in Dr. Wachter's lab. We learned that it is absolutely imperative to have a steady hand when placing the mix of buffer, dye, and polymerase into a gel well due to its small size ( at least it was for me...since I had to redo it!). Fortunately, we actually had the dye. The researcher informed us that sometimes this has to be done with colorless solution so the level of difficulty is increased. We waited for an hour, added another dye, and mixed it for 15 minutes and then saw the sample DNA along with the segment we had replicated underneath UV-C light- picture forthcoming here.

We also listened to the research of Dr. Wachter and I learned the real-world applications of her work. Essentially, if you can input GFP- green flourescent protein- into a nematode (it is originally from a jellyfish) and target nerve cells to flouresce specifically when they are firing, you can use this particular protein as a "biosensor." Since, CFTR malfunction is the main cause of Cystic Fibrosis, we could then use a GFP clone to follow the misfunction via flourescence lighting up a specific piece of information such as chloride transport in this case.

Lastly, GFP takes aboput an hour to turn green but an hour may be too long with live cells so Nam- a researcher- is currently working on the development of GFP variants that mature- "turn green" - more rapidly and this is done by creating a DNA library to look for the ones that flouresce quicker- similar to artificial selection in dogs- i.e. breeding selectively for specific morphological or behavioral traits.

Question to ponder:

Can we improve upon nature?

Certainly depends upon specific conditions.

Today, we met with Dr. Wachter who re-introduced me to PCR analysis. I was very excited to be performing this relatively simple procedure since I have been teaching it for many years but have never had the opportunity to actually perform such procedure. It was just as simple as I thought, except for the minute quantities we were dealing with. Apparently, many of the materials were simply purchased from larger lab corporations. Then, we had the opportunity to see how agarose gel was mixed. I have to admit that I felt much more comfortable in this type of lab. Later, we changed concentrations several times to get the correct amount of crystals for seeding for PS I. Then, we had to work with a dialysis tube and capillary with a very small "loop" and we had to place it in a miniscule tube without touching the side in a cold room at 4 degrees Celsius! Fun, adventurous activity!

7/9/07

Today, we tried a dialysis technique to crystallize PS I which required the use of many different equations as well as a series of rather complicated steps such as preparation of probes,detremination of chrolophyll concentration, reactor assembly. We were able to see "fast and immediate crystallization," use the spectrometer again, and deliberately denature the protein. We learned the secret of crystallization of PSI- sometimes you need to use a lower salt concentration. We are "recrystallizing" essentially for PS I or will be microseeding tomorrow and working with Dr. Wachter for PCR.

This work was painstaking and complex to me. I am excited to see what will happen due to our groups' work.