1-28-2010: Last day of the internship

Today was a really good day. It was pretty relaxed, all I did was dry samples that Dr. Nuss will run on a western blott next week. I also talked to Dr. Brown about my experience in the lab and he helped kind of connect the small experiments we have been doing into the whole grand scheme of all the research that has been done on invertebrate insulin-like peptides. During my off time I worked on my paper some more and started putting photos together for the presentation.

Tomorrow I will be shadowing Dr. Harper, a general surgeon.

1-28-2010: Results Day

I woke up this morning and realized that I did not post a blog yesterday... But we had a great day yesterday because we got really positive results back. The western blot that we have been running for the past three days with bloodfed vs. nonbloodfed showed phosphorylated tyrosine in the bloodfed mosquitoes and it was not seen in the non-bloodfed mosquitoes which is what was expected.

It was also good because I took a lot of pictures for my presentation next week and everyone thought that was fun. I was also able to take some pretty cool pictures of dissections through the microscope lense which I didn't think was going to work. This was my favorite of the dissections. From the left to right is the head, thorax, and abdomen of an Aedes aegypti mosquito. We seperate these parts in order to test the presence of peptides in certain areas of the mosquito.

1-26-10: paper feedback

Today I showed Dr. Nuss some parts of my paper that I had finished. I had written the western blott procedure for the materials section with a lot of my own notes and steps on how to do the procedure, but he said for a more scientific paper you only need to put the ingredients for the buffers, temperatures and more conditions than actual instruction on how to do the experiment. I also did a lot of dissections - 250 Aedes aegypti heads. We also did added the primary antibody to our blott and it is being washed on a rocker in the cold room right now.

I am working on my paper some more tonight and tomorrow we will be finishing the last western blott I will be here for and getting our results.

1-25-2010: Western blott and dissections

Today I basically did the same thing I have been doing the past two weeks. We started another western blott today with our bloodfed dissected mosquitoes and the non-bloodfed mosquitos in order to compare the amount of phosphorylated insulin receptor in the two groups. Today was more exciting because I did a lot of the western blott on my own. I set up and ran the gel and prepared the nitrocellulose paper and most of the western blott sandwich for running. I also took the gel and the paper out, which is one of the hardest parts of the procedure because the gel can stick to the paper and if it dries then you can't read it and it's important not to tear the gel and the paper. I did mess up a little and couldn't get a small piece of the gel off, but luckily it was over a part of the paper that didn't matter too much and it was ok that it didn't come off there. I also did a lot of dissections for the next western blott we will run that I probably will not be able to see.

Over the weekend I got my TB test and background check so that I can shadow in the OR on Friday and I will be shadowing a general surgeon, Dr. Harper. I also worked on my paper a bit over the weekend and Dr. Nuss said that he would read it tomorrow to make sure it's accurate and organized well.

1-22-2010: Shadowing Dr. Larrimore

Today I shadowed Dr. Larrimore who is an OBGYN in Covington, GA. I arrived at her office a little before 9:00 because she was doing a procedure at 9. However, her patient did not sign the consent form to allow me to view the procedure, but I was able to follow a midwife who works in her practice. The midwife did an ultrasound on a woman who is pregnant with triplets and showed me how to read an ultrasound. That was really cool because Dr. Larrimore said that she has been in practice since 1996 and this was only the second case she has had in which a woman was going to have triplets. The midwife also checked the heartrate of the baby of a woman who was about 15 weeks pregnant. By the time we had done that, Dr. Larrimore had finished the procedure. She had removed a benign tumor from a young pregnant woman and showed it to me.

Then we went to the hospital and saw three patients. One woman had just had twins and was in a hospital bed recovering. She was having a sharp pain near her right hip and Dr. Larrimore said that she may have an infection and had a nurse do a urine test to check for infection. The second woman we saw was having contractions and about to have a baby. We just went in to check on her and see if she needed any pain medication. The last woman we saw had just had a baby and was leaving the hospital that day. We also went to the area of the hospital where they keep the babies right after they are born. I talked with the nurses that work there and they told me all about their job and how peaceful it is unless they have a baby that is going through drug withdrawl because they are always irritable from being weened off of whatever drug their moms were on while they were pregnant. We also met and spoke with a radiologist who had done an X-ray on one of Dr. Larrimore's patients. Dr. Larrimore said that they usually only work halfdays on Fridays and I left during lunchtime around 11:30.

Now, I am about to write in my shadowing log about what I did today and I am going to write up a rough draft of the western blotting procedure we have been doing in the lab for my paper.

1-21-2010: More dissections...

Today I and Dr. Nuss did a lot of dissections, but these were different from the previous two types of dissections we have done so far. For these dissections, we isolated the midgut of the mosquito and will test on a western blott to see if a phosphorylated insulin receptor is present in the midgut of bloodfed mosquitos. The results will be compared to the peptides we find in non-bloodfed mosquitos. We expect to see a lot of phosphorylated insulin receptor in bloodfed mosquitos because our hypothesis is that bloodfeeding starts the insulin pathway. We believe that proteins from the blood interact with a receptor in the midgut (which may or may not be present... we will see) which then sends a signal to the brain and causes neurohormones to be released and sent to the ovaries of the mosquito which signals egg production and ultimately controls the reproduction of the mosquito. So, in order to isolate the midgut in order to test for the phosphorylated receptor, we put about 100 mosquitos on ice in order to anethesize them and then decapitated them so that we could pull the midgut out of the mosquito from the tail (part of the midgut is attached to the head of the mosquito which is the reason for the decapitation). By pulling on the bottom tip of the abdomen of the mosquito, the outer layer rips and the attached ovaries, digestive tract, midgut, and salivary glands all come out in one connected string of organs. We then seperate the rest of the organs from the midgut and then put the midgut into solution (made up of ingredients that preserve the midgut and inhibit proteases from degrading our target peptide). We had to isolate the midgut from 40 bloodfed and 40 non-bloodfed mosquitos, so we did that and then prepared the extraction solution. After setting the midguts into dissection solution, centrifuged them and then resuspended them into homogeniztion solution (and I am not sure what the function of this solution is). We then ground up the pellet into the solution in order to release more of the target peptide. We then stored our samples in the -80 freezer.

In the midst of doing dissections, I also prepared the western blott for the primary antibody wash. I put the antibodies on and the nitrocellulose paper is soaking in the antibodies overnight.

Tomorrow I will not be shadowing Dr. Harper because I found out that I have to have a background check and a TB test in order to shadow in the operating room. However, I will be shadowing an OBGYN, Dr. Larrimore, tomorrow.

1-20-10: Gel Day

Today we started a new western blott with the samples we prepared yesterday. This morning, we first prepared the samples by adding ethanol to rehydrate the peptides and sample buffer. We then vortexed them to resuspend the proteins, poked holes in the top so that pressure would not build up in the tubes during heating and then put them in the incubator at 55 degrees Celsius. After the samples were prepared we put them on ice and loaded them onto the gel to be electrophoresed. We ran the current through the gel for about three and a half hours. After that, we prepared the western blott sandwich by cutting the blotting and nitrocellulose paper and soaking them in 10% methanol. Then we removed the gel and put the sponges, blotting paper, nitrocellulose paper, and gel into the sandwich. The proteins are being transferred from the gel to the paper now and will be ready around 4:05. After the paper is ready, we will remove it from the sandwich and set it out on our lab space to dry overnight.

Dr. Brown gave me some extra textbooks today on endocrinology that should help with the primary literature I have been reading. I have been reading one of the books during some of the down time today.

I also spoke to my dad last night and he said that he had arranged for me to shadow Dr. Harper, general surgeon on Friday.

Tomorrow we will label the paper and do the primary and secondary washes over our western blott.

1-19-10: Sample Preparation

Today we have been working on preparing samples that we will run on our next western blott. On Thursday, the workers in our lab and I dissected mosquitos by seperating the head, thorax and abdomen. Dr. Nuss mixed different solutions that should extract different proteins from the mosquito parts into the solution. On our last western blott, we did not see any proteins from Aedes aegypti in our results, and Dr. Nuss wanted to try a couple of new extraction solutions that might cause the proteins we are trying to identify to go into our sample better. We took the solutions he prepared and centrifuged and dried them. Once they are all dried, we are going to resuspend them into a smaller amount of alcohol so that the protein content of the solution will be more concentrated than it was in the extraction solution. Then tomorrow we will begin running the samples on a gel. While the samples have been drying, I have been dissecting more mosquitos. Another postdoctorate associate in the lab is doing an experiment to isolate the mosquito insulin receptor (MIR) in the ovaries of the mosquito and then add it to synthesized ILP3 in order to test the affinity of ILP3 with the receptor. The dissection I did this morning was a little bit different; because the ovary needed to be isolated, we dissected by pulling the lower tip of the abdoment from the rest of the mosquito, which then caused the ovaries and intestinal tract to be pulled out of the mosquito abdomen. We then placed each pair of ovaries into a buffer so that he will be able to prepare a sample from the collections.

Last week when we photographed our results, I saved the pictures of our results onto my flashdrive in order to post them onto the blog, but I was unable to do so I think because the images are saved as .tif files. So, Dr. Nuss said that I would be able to label each of the lanes with which sample it was and change the tif file to a jpeg file with photoshop. So I installed photoshop onto one of our lab computers and am going to try to modify and post a pictures of last week's results.

This image is a photograph of the results of our first western blott from last week. Before adding the antibodies to the proteins on the paper, we cut the paper into three different segments so that we could test 3 different types of antibodies on our samples. The middle piece of paper was washed with a purified primary antibody while the other two side pieces of paper were washed with a raw primary antibody. The piece of paper washed with purified antibody did not show on the first photograph (30 second exposure) because there is no binding going on in the background. Antibodies are made by injecting the target proteins into an animal (in this case a goat), and in response, the goat's immune system makes antibodies. The raw primary antibodies include antibodies for other things that the goat encountered and are not seperated from our target antibody for ILP3. For that reason, the background of the paper washed with the raw primary antibodies are darker due to binding from the other antibodies in the raw solution.

Before we ran the gel, we mixed our samples into two different types of buffer (NuSep and Reducing). The reducing buffer functions to break the disulfide bonds that connect the A chain and B chain of the ILP. The NuSep does not do that and it is harder to compare the sizes of the bigger peptide with the A chain and B chain together with the ILP3 A chain standard and ILP3 B chain standard. The size is marked by the ladders that are in the middle of the three sheets of paper.

The lefthand segment of the paper was ILP3 A chain and B chain standards as well as Aedes aegypti head extractions (done via dissection, grinding and mixing with protein extraction solutions). If you look at the bands on the paper, all of the ILP3 chain standards showed up, which is good because Dr. Nuss had done a previous experiment in which they did not show. However, we did not see any bands for the Ae. aegypti heads. For that reason, today we prepared Ae. aegypti heads in different extraction solutions to run on the gel in hopes that we will be able to detect this ILP3.

On the segment of paper to the right we tested ILP134 standards and Anopholes stephensi heads. The ILP134 standard is made up of different segments of conserved ILP1, 3, and 4, and the antibodies should recognize any one of the three ILPs. Our lab has hypothosized that the function of ILPs 1, 3, and 4 is somewhat redundant and that is why we tested for all three of the ILPs. On this blott, we did get a band for the mosquito heads, but it was bigger than what was to be expected. If you compare the size to the standards, you can see the difference in the sizes.

Also, we tested the standards again by dotting the standards onto the nitrocellulose paper prior to the primary and secondary antibody washes. You can see this best on the segment of paper to the left in the bottom lefthand corner. We did this as a positive control because if there was anything wrong with the process before the antibody washes, then we would have seen the ILP3 dotted standards, but not the standards that went through electrophoresis on the gel and onto the paper. There is a small dot which is the B chain ILP3 standard. The different standards and bands look better or worse depending on the exposure time as well.

To finish the day, I am going to continue to help Animesh with ovary dissections.

1-18-10: MLK Day

Today I did not go into the lab because UGA was closed for MLK Day. But I did read a couple of articles to get some information for the paper requirement, start a log for the time I spend shadowing physicians, and make the rough draft for my presentation. The papers I read were "Signaling and Function of Insulin-Like Peptides in Insects" and the author is Dr. Mark Brown (whose lab I am working in) and one of his undergraduate students, Qi Wu. Dr. Brown gave me this article because it is a review article of what is known of insulin like peptides in several different species of insects. It was helpful because it improved my understanding of insulin like peptides, but I will not be able to use much of the information for my paper because I really want my paper to focus on what is known about ILPs in mosquitos and much of the information in this paper was about ILPs in Drosophila melanogaster. I also read "An insulin-like peptide regulates egg maturation and metabolism in the mosquito Aedes aegypti." As the title says, the paper was about what the known effects of ILPs are on reproductive and metabolic function in the A. aegypti mosquito. The paper had a nice description of the structure of an ILP, some basic information about the life cycle of the mosquito, and some background information on human ILPs - all points that I will include in my paper. The study showed that ILP3 in the mosquito was most similar to human insulin and was also found to be present in the mosquito brain. It also showed that ILP3 stimulates egg maturation in mosquitos. They found this by decapitating mosquitos and then injecting them with synthetic ILP3. The ovaries of the decapitated mosquitos then began to secret ecdysteroid hormones which caused that fat body to secrete yolk proteins (which were derived from the blood meal) in order to form the eggs. It also identified the receptor for ILP3 (mosquito insulin receptor - MIR) and showed that ILP3 without the receptor does not work. They did this by injecting double stranded RNA of the gene that controls the MIR so that the RNA would be degraded and the gene would not be expressed (therefore causing no insulin receptor to be present). In the absence of MIR, with the injection of ILP3 no reaction occurred. These two paper will be very useful for when I begin to write my paper, and I am glad I had this day to read and take notes on them because I have not had much of an opportunity to read so far.

Over the weekend I bought a notebook that will serve as my log for all of the times that I shadow a physician. I recorded my time with Dr. Manning and wrote a few notes about what I thought of the visit in it. I also made a rough outline of my presentation.

1-15-10: Shadowing Dr. Michael Manning

Today I shadowed Dr. Manning who is a family practicioner in Conyers, GA. I absolutely loved shadowing him and I really love his job. I am really glad that I have been able to compare researching in a lab with shadowing a physician this week.

One thing that I don't like about researching in a lab (at least the one lab that I have experienced) is that it's not a very social job. There aren't too many people in our lab, and the people that are there generally don't collaborate too much. Most of the people work on the same general theme but they all do their own projects and independent research. In comparison, Dr. Manning's office was much more of a social atmosphere. I really loved talking to all of the patients, nurses and doctors in his practice. Just that aspect of his job was a big improvement in the nature of the work and more suitable to my personality.

One reason I sought out Dr. Brown's lab to shadow is because I am very interested in malaria and research that could possibly improve the condition of people with malaria or prevent the transmission of malaria from the mosquito to people. One thing that I don't like about working on the research is that it is a very distant way of bringing relief to people. The mentality of the research is that we are doing this research because there is a correlation with insulin and mosquito lifespan and transmission of malaria which could help to decrease the number of people affected, but first, we need to learn more about ILPs and the insulin pathway in mosquitos which is a very indirect way of solving a problem (and I am so I glad that there are people who are willing to do all of the research and work for medical advancements, but I don't think that I want to do that). I really liked the patient/doctor relationship between Dr. Manning and his patients and the direct way that he was able to improve their conditions and help them to live better and healthier.

One thing that has kind of kept me from definitely wanting to go to medical school is the fear that I would go through a lot of school, have a specialty and then eventually have an incredibly repetetive job and not really learn anything new too often. Dr. Manning told me that because he has such a general practice, he has to know about so many different conditions and problems in every part of his patients' bodies. He also said that because of technology there are always advancements and new treatments that he has to continually read about and learn about in order to provide his patients with the best care. That is one aspect of his job that he likes and something that I really liked about his field as well.

Today I arrived at his office around 8:30 and saw patients with him until about 12 and we had lunch and then went to see a patient of his in the hospital until 2. Then we came back and saw patients for another hour or two. He stayed later but I left around 3:30. I really enjoyed the experience and it has made me much more excited and enthusiastic about potentially applying to medical school.

01-14-10: Mosquito dissection day

Today I dissected 144 mosquitos... For our next Western blott we will be testing to see if phosphorolated (active) insulin receptor is present in the Anopholes stephensi mosquito. We will also be testing to see where the receptor is present (the head, thorax and abdomen.) So... today, Dr. Nuss and I collected about 150 mosquitos and anethesized them by putting them on ice. Then we removed the legs and wings and then seperated the head, thorax, and abdomen. I did that basically all day. Then we took what we had collected and grinded it up with a pestel and kept them in a -20 refridgerator. Tomorrow I will be shadowing Dr. Manning.

1-13-10: Results today!

First, I am going to lay out the Western Blott procedure because I do not think I have fully understood it until today and could completely explain it in a simple way:
1. First, you prepare your protein samples (our samples are controls that definitely contain insulin like peptides and mosquito body parts that we don't know if they contain insulin like peptides or not).
2. Load the samples into a gel and electrophorese in order to seperate the proteins into different sizes. Proteins with a higher molecular weight appear higher on the gel and proteins with a lower molecular weight appear lower on the gel. We also load markers which indicate the size of the proteins that show up in our sample.
3. Transfer the proteins from the gel to nitrocellulose paper. This is done by placing the paper next to the gel and running another electric current through the gel in order to pull the negatively charged proteins from the gel to the positive pole and onto the nitrocellulose paper.
4. When the proteins are on the nitrocellulose paper, they are not visible. To make them visible, we add antibodies that are specific to bind with the ILPs.
5. We then add secondary antibodies to the proteins and the antibodies that is specific to bind to the antibodies. When the secondary antibodies bind to the antibodies, a reaction occurs, a color is produced, and you know whether or not ILPs are present in the mosquito heads.

Today, Dr. Nuss and I finished our first Western Blott. We left the protein samples on the nitrocellulose paper to soak in the primary anitbodies overnight last night. When we came in this morning, we did 3 washes for twenty minutes each with the TBS-T (tris saline buffer with tween 20). Then, we added the secondary antibody to our proteins and allowed that to soak for about four hours. And then we were able to photograph our samples and view our results. Before I started working this week, Dr. Nuss ran a Western Blott with chain A standard, chain B standard, mosquito heads, thoraxes and abdomins. The bioactive form of insulin is made up of an A chain and B chain connected by a disulfide bonds. The chain A and chain B standards are the segments of insulin, and should definitely show up on the Western Blott. One of the first times Dr. Nuss ran the procedure, he ran it with these standards, but for some reason the standards did not show up in the results when he ran it. The Western Blott we have been running the past three days included chain A standards, chain B standards, and mosquito heads. In our results, we were able to see the standards, so the procedure worked, but there were no bands for the mosquito heads. Dr. Nuss said that he thought it may have been a problem when making the mosquito extract.

We also started our second Western Blott by preparing our samples and running them on a gel. Then we transferred the proteins from the gel to the nitrocellulose paper and the paper is on the lab counter drying.

Tomorrow I am going to finish the second Western Blott that we started today and dissect some mosquitos because we are going to increase the number of mosquito heads in our next Western Blott to see if increasing the amount of mosquito juice will give us a band on our next Western Blott.

I also spoke with my dad (who is a gastroenterologist) and he has arranged for me to shadow one of his friends, Dr. Manning - a family practicioner, this Friday.

1-12-10: Mosquito feeding day...

Today Dr. Nuss and I started the second stage of the Western Blotting procedure which is called the immunoblot stage. During this stage, we took our dried nitrocellulose paper with the blots of our insulin like peptides and first blotted what are called standard ILPs onto the paper as a positive control. The standards are solutions that definitely contain ILPs and we blot them onto the nitrocellulose paper as a positive control because if the paper is somehow defective then the ILPs do not show up and we can know that if we don't see anything from our experiment that it is because of the paper and not another problem in the procedure. We then cut each piece of nitrocellulose paper into three segments according to which wash each sample needed to receive. We then soaked each of the pieces of nitrocellulose paper in TBS-T (triss buffer saline with tween 20) for a couple of minutes.

In the immunoblot procedure, there are two main washes that need to be performed: an antibody wash and a wash that contains a catalyst such as an enzyme. The first wash is the antibody wash because the antibodies in the liquid bind to the proteins on the membrane of the nitrocellulose paper. The second wash contains the catalyst and this portion of the procedure is done because the enzyme binds to the antibody and then a reaction occurs which produces a color and allows for the protein to be visible. We performed the first of the two washes by preparing a block solution from ECL and TBS-T and drained the TBS-T from the paper and poured our newly made block solution onto the paper and put it in a rocker in the cold room. After about two hours we took our papers from the coldroom and injected antibodies into each of the dishes containing the block solution and paper. Now, the samples with the antibodies are on the rocker in the cold room and we will finally get our results tomorrow.

Then we made up some new TBS buffer by mixing NACL, HCL, Tris base and deionized water. It took a while to make the buffer because the HCL that we were using was highly concentrated and dangerous to handle. We only handled a small amount at a time (about 10 mL) and then safely disposed of the extra that we had left over.

Then I labeled 30 small tubes because tomorrow wer are going to alloquat a large amount of an antibody solution into the smaller tubes so we can put the solution into the -80 degree refrigerator so that bacteria does not begin to grow in the solution. Dr. Nuss explained that it is important to put the solution into small amounts because if you freeze and thaw a solution containing proteins several times then the proteins begin to degrade.

Before I went to lunch, I went into the mosquito room because another person working in our lab needed to bloodfeed the mosquitos so that they would mature and be able to produce eggs. He had anethesized a rat and put it on a screen for the mosquitos to feed on it and it was kind of neat to watch. He is doing an experiment that involves injecting different solutions into the mosquito that will hopefully stop the genes controlling insulin like peptides from being expressed. He let me inject one of the mosquitos using a small needle and syringe and it was really tedious, but it was kind of cool because first he froze them so they could pick them up and handle them, injected them and then put them in a container and right after the injection they would start flying around like nothing happened.

Tomorrow we will get our results and I think start a second western blot.

01-11-10: First stage of Western Blotting technique

The main project that Dr. Nuss and I will be working on is where insulin like peptides are located in the body of the mosquito. At the meeting last Thursday in the Holcomb room the question of whether the insulin like peptides (ILPs) in the mosquito are produced by the mosquito or if they are from the insulin taken up in the blood meal. I asked Dr. Nuss this question and he said that there are several ILPs, ILPs 1-5, and that they are all endogenous, meaning that they are all produced by the mosquito. Dr. Nuss also explained that part of the mosquito's immune defense system is to release free radicals in order to kill outside pathogens that may be harmful to the mosquito. One of the functions of insulin like peptides is to suppress the mechanisms that control the free radicals. So, when high amounts of insulin are present, free radical production is higher because there are fewer active mechanisms to control the put out of the radicals. When the free radicals are released within the mosquito more often, there is more damage done to the mosquito, and its lifespan is shortened. Although Dr. Nuss and I are not working on this aspect of the research, the lab is doing research to find out if a blood meal containing high amounts of insulin will ultimately shorten the life of the mosquito because of these mechanics. Dr. Nuss and I are working to learn more about insulin like peptides in the mosquito species Anopholes stehpensi and we are starting by doing experiments to find out where insulin like peptides are located in the body of the mosquito.

One type of experiment that we will be doing to identify ILPs in the mosquito is a technique called Western Blotting. Last week, before we were able to start experimenting today, Dr. Nuss had to do some preparations to run the Western blot. One thing that he did before we could start the experiment today was to disect about 500 mosquitos by seperating the heads from the body, grinding them up, centrifuging them and collecting the supernatant to run for the Western Blot. We are doing this experiment in order to identify whether or not the ILPs are located in the head of the mosquito. Today, there were three main parts of the first stage that we ran: preparing the protein samples, running the samples on a gel, and then transferring the proteins from the gel to nitrocellulose paper. We had 16 samples to run on our gel, and they were all either samples of mosquito heads, chain A of insulin or chain B of insulin. Insulin is a hormone made of of chains B, C, and A, but chain C is cleaved out of the active form of insulin and chains B and A are connected by disulfide bonds. So the samples with chain A and chain B are just insulin. We mixed the chains B and A, and the mosquito heads with different buffers (sample buffer and reducing buffer - which serves to bind with proteins that may be "sticky"and bind with anything, such as the proteins we are trying to detect) in order to make our 16 samples. Once our samples were made, we loaded them into a gel and did electrophoresis for about 3 and 1/2 hours. During that time I went to part of a seminar on termites and how they could be a valuable energy source because they could be used as a high calorie food or to produce high amounts of methane gas. Then I went to lunch with the program director, Dr. Marianne Robinette. After that, Dr. Nuss showed me how to set up the next part of the Western blot. During the remainder of the time to wait for the gel, I read a primary literature article that relates to the research we are doing. Once the gel was ready, Dr. Nuss showed me how to prepare the Western Blot "sandwich". We prepared two pieces of nitrocellulose paper on two sponges and then put our gel in between the two pieces of paper and into a machine to transfer the proteins on our gel to the nitrocellulose paper. The transfer to the nitrocellulose paper took one hour and I read part of another literature article. Then we were able to look at the nitrocellulose paper and our markers were visible. Tomorrow we will need to do a couple of washes over the paper that will cause the proteins to change color and be visible in order to see if they transferred to the paper. That was all we did for today and tomorrow we will be finishing up the Western blot.

Jan 08 2010

Last Tuesday I went and visited Dr. Nuss and he gave me some reading that I could do over the weekend. Today, I went to Target and bought a binder and notebook so that I can outline from the Molecular Biology text book that Dr. Nuss gave me. I also organized teh papers he gave me and the papers that Linda Powers passed out last night at the meeting.

UGA Internship - Meeting in Holcomb Room

Today we had a meeting and each member of the research in the sciences and humanitites interims had a chance to explain the type of research they will be doing. Last Tuesday, I went and visited the lab I will be working in and spoke with Dr. Mark Brown and Dr. Andrew Nuss, the professors I will be working most closely with. Dr. Nuss gave me a brief outline of the Western Blotting technique which I will be using to identify insulin like peptides in different areas of the mosquito. I will also be sitting in on classes every Thursday and shadowing a physician every Friday.