Blogs from June 26th-29th

Jannette O. Nuestro
Blogs from June 26th -29th
Photosynthesis Fromme
May I express my apologies for not having been able to post my blogs online. I wasn’t connected to the internet until today. However, I have been keeping and writing my reflections for each day regarding my experience in the research lab with Dr. Fromme.
June 26th Tuesday
We started today with our lab with the help of one of Dr. Fromme’s graduate students named Yana. She took us for a tour of the different lab areas where they perform the crystallization of protein. The plan for the day was to prepare the chemicals needed for the crystallization of protein called lysozyme in plants. Yana was very helpful, willing and patient in reviewing and reteaching us the chemical equations necessary to arrive at the desired units of measurements for the chemicals needed for the lab. It was an experience that took me back in time many, many years ago when I had chemistry in class. I feel like a student again having to jog my memory of solving chemical equations. When the chemicals were prepared, Dr. Fromme took us into her office for a powerpoint presentation on photosynthesis. The presentation gave us an introduction to the research that she has worked on and gave us an overview of the different experiments that will be performed in the next few days.
June 27th Wednesday
Today, we worked in the lab with Dr. Fromme in crystallizing lysozyme in plants. The method we used today is called batch crystallization, where the lysozymes or enzyme will be crystallized inside the thin glass capillaries. The chemicals used where lysozyme, sodium chloride, acetic acid and sodium acetate – these were the chemicals that were peepared yesterday. We used different materials in the lab, such as micropippetor , plastic vials, capillaries, permanent markers and tape. I had to mention permanent markers and tape to emphasize its importance along with the micropipettors because these items are as we found out are just as important as in keeping track of our experiment. Everyone of us in the group had a nice turn in working in the experiment using the pipettors in obtaining accuracy of the mixture in chemicals. And we all found out that just as our students in the classroom, we made our mistakes and had to repeat our mixture of chemicals at least two times. We were just glad that one of us noticed the mistakes and we were able to correct it.
June 28th Thursday
Today, we looked at the capillaries form the batch crystallization hat we have prepared from
yesterday.We made two groups of cru=ytallization with varyig amounts of lysozme. We labeled the first
group with 100 mg./ml. of lysozyme as batch A and the second group with 50 mg./ml. of lysozyme as
batch B. Both groups had the same amount and
mixture of chemicals of sodium acetate (NaAc), sodium chloride (NaCl)and water (H2O). Different
capillaries have varying amounts of proteins that have cruystallized depending on the amount of
repetitions and mistakes the group made yesterday. But, like any science experiments, there is always
room for errors J. In addition to seeing the results of our experiment from the previous day, we also
started with two more experiments. The goals of these experiments was to use different methods of
crystallizing protein. The first one which was much easier to prepare was called free interface-diffusion
The protein solution used for the experiments use 50 mg/ml lysozyme and the same amount of sodium
chloride with sodium acetate as the precipitant. The second experiment is called vapordiffusion. This
experiment stores the chemicals in a reservoir
with mixtures of the same chemicals, sodium acetate, sodium chloride and water. On a
separate glass slide is the protein solution. This will be stored overnight and be observed the
following day.

June 29th Friday
Today, we had the opportunity to sit with the graduate students under Dr. Fromme and to listen to one of the student’s powerpoint presentation on his experiment. His presentation was on the expression of protein in Spinach in E.coli bacteria. It was a very interesting experience to watch the grad students listen and offer suggestions on the presenters experiment. After this experience, we went back to the lab to look at the results and take pictures of our experiments from thursday. Once again, with pride and enjoyment, we were successful and have seen beautiful crystals formed under the dissecting scopes. Dr. Fromme explained the differences in the amount and sizes of the crystals formed and showed us how to take pictures of the crystals. Pictures that we can take in the classroom to show our students and post in our wiki J. The whole week was a wonderful and productive experience with Dr. Fromme . She had been very professional, dedicated, willing to teach and help and encourage us to experience for ourselves being in the research lab by actually doing the experiments and using the materials in the lab. She had been very accommodating, very well organized and prepared us well everyday for each experiment.