Spring Break Plans

Spring Break began yesterday, and I have a week off from school. I am so happy that I get to forget about school for a week :).

Today, as I was driving home from Fayetteville to Springfield, the weather began to change for the worse. When I started my drive, it was raining, but by the time I reached Joplin, the rain turned into snow. (eeep!) So I pulled over for a while waiting for the snow to pass, as I did not feel safe driving on I-44 in possibly icy/snowy conditions. But, luckily, after about 15 minutes or so, the heavy snow became lighter snow, and then eventually turned into rain once again. So I didn't really have to drive in the snow, just in the rain ...which is fine.

The only real plans I have other than just relaxing and being with family is to volunteer at the Kitchen Clinic. It's a free medical clinic in Springfield, and they are always in need of volunteers. I plan on working on Monday, Tuesday, and Wednesday. It's a pretty rewarding experience.

Oh, and another thing I'm doing this Break is going to my brother's friend's wedding. He's been a family friend since I was very little, and so he has invited my entire family to attend his wedding. (My sister can't attend because she is working overseas in S. Korea). It should be fun though, and it will also be the 3rd wedding I've ever been to.

Then it's back to school until May.

Busy Week

This week is somewhat busy, I have two tests tomorrow (Wednesday). One is in physics 2, and the other is in music literature. Obviously, the physics test will be difficult, but the music literature is not so easy either. Mainly due to the "listening" part of the test. Each test, we have sixteen pieces of "classical music" from four or five different composers (usually), and we must be able to identify each piece only through a 30 second random sample from their corresponding piece of music. This part can be tedious, as the symphonies from Beethoven, for example, can be quite lengthy, and the sample given during the test may not be very representative of that particular piece, so it may be hard to recognize. I can distinguish between most of the songs, but there are a few that I still confuse at times.

This week I won't be doing any research, as my professor is very busy before Spring Break. This is good for me as well though since I have tests this week.

There is only one Kaplan prep class left, which is tomorrow. Then, every weekend after that we are scheduled/encouraged to take a practice exam mcat. I am still not as prepared as I would like, but since I have three more months until I take the exam, I feel confident that I will have sufficient time to prepare. Especially the last month before the exam, where I will not have school work to keep me busy.

I am ready for Spring Break!

The Return of the Droop-Monster

My puglet, Droopy, can now speak on command. It's the cutest trick ever. Since he is still reluctant to bark, he will make a pathetic/cute half-bark at first and then follow with a big dog bark. Maybe I will add a video of him later. The way Droopy learned this trick is kinda strange. Pancho hung a new picture on the wall, and for some reason Droopy hated it and began to bark at it. We said "good speak" and gave him a treat. It didn't take long for Droopy to bark if we looked or pointed at the picture, and eventually he barked without the use of the picture, only with verbal cues. He also has learned high five recently, but that trick is very similar to "shake" so it's not as impressive. He can also "stand up" for quite a long time.

I plan on taking him on a walk today after my classes, because he loves going on walks. He does need a lot of training when it comes to walking, though, as he tends to pull. I avoid him pulling by running with him at the beginining of our walk so that he releases a lot of his energy. After he gets tired of running (max 2 minutes), then he will be a good puppy and walk. I am teaching him "heel", and he seems to be learning it. Sometimes he will actually respond correctly to the command, but other times he is confused by it. (I can tell when he is confused because he flexes his ears really far back so that the wrinkles on his forehead are stretched and smooth, and looks at me with puppy eyes like he's in trouble). Nevertheless, he doesn't let the commands get in the way of a good time.

He has an adorable little gallop and slobbery mouth when we walk, and he is very attentive to me. He tends to look up at me every now and then, smiling with his curly tail wagging side to side. But, because he is a brachycephalic dog, he runs the risk of being easily overheated, so i bring some water to splash on his face/for him to drink, and keep the walks short. He is always exhausted after the walks, though, and has to take a loooonnnggg nap on my lap afterward.

Step by Step Instructions for PCR

Step by Step Instructions for PCR
1. Prepare your chemicals/reagents and Work Area
Take Taq, dntp, primers (2), buffer solution, MgCl2, from the PCR box in the fridge. Mix the primers thoroughly.

Then, put the water on the heater to kill any bacteria or foreign agents that could be inhabiting the container.

Remove DNA samples that are needed from the fridge and arrange in tray according to the order on the chart (unique for each PCR reaction).

Next, count the number of different DNA's that you will be using (usually around 15), then add one more for the control, and then add one more to provide for possible errors (such as pipetting). 15+1 control+1 for error= number 17.

2. Mixing Reagents into a Medium Test Tube Carefully
Using this number, refer to the chart posted in the lab, and add the required amounts of each of the PCR reagents listed above to a medium sized test tube. (the water can now be removed from the heater). Make sure to mix the Taq (gently by hand) before adding it to the mixture.

After you have added the correct amounts of each of the reagents, you place the test tube into the centrifuge for only 5-6 seconds to make sure everything is mixed together.

Then, according to the number of DNAs (+1 for control), you will use that number of tiny, tiny test tubes. Today I had 15 DNA samples, 1 control= number 16. (The extra one that I used for error remains in the medium sized test tube, tossed away in the end). Then, I assigned each DNA to a number, and wrote one number on each of the test tubes, so that I can know which DNA is in each tube. (denoted the control with a - sign).

3. Fill each tiny tube with mixture of concoction and DNA
Next, pipet 21 microliters of the concoction from the medium sized test tube into each of the tiny tubes. (Careful to avoid contamination). Next, pipet 4 microliters of the particular DNA sample that correlates to the number assignment on each of the tubes. Makes sure to pipet both the concoction and the DNA into the tiny tube in such a way that the liquid is at the very bottom of the tube. If this is not done, then the tiny tubes must be put into a small centrifuge to spin all of the liquid to the bottom of the tube.

4. Heating Process
Now you are almost done. You put all of your labeled tiny test tubes into a machine that heats the samples in order for the entire process to begin. (denature the protein, split the double strand into two single strands... refer to previous post for more detail).

This heater runs for a couple hours (time depends on the primers used) in different cycles. The machine is already preprogrammed, so I don't know the exact times for each cycle.

After the heating process is done, you put your samples into the fridge to wait for the next step of the process: gel electrophoresis.

Lab work

Tuesday was my first day working in the lab with Dr. Dowling. What I did was PCR, which stands for Polymerase Chain Reaction. Basically, the DNA sample is denatured by a high temperature (for example) and due to this, the DNA splits into two strands. Then, on each strand, PCR basically rebuilds a new base pairing strand to complement the existing strand. So, each time you run the reaction, you get double the amount of DNA which you started with. (Started with one DNA, split it into two strands, and built upon each of those strands for each of them to become a full DNA double helix form again. Thus, you get two identical strands of DNA from one DNA. So, as you can see, the number of DNA grows exponentially. Anyway, specifically what I did in lab on Tuesday was pipetting different primers, buffers, Taq, and DNA, to form a concoction. This concoction was then pipetted in 25 microliter volumes into tiny tiny test tubes. This process took a while, but is not difficult. It requires attention to detail, steady hands, and meticulousness. You have to be sure not to introduce any foreign agents to the mixture, and must maintain a clean environment. If the tip of the pipeter touches anything save for the mixture or the inside of the test tube, then you must discard that pipetor tip and begin again. Cross contamination must also be avoided. I had to discard the tip a few times because I accidentally touched it to the outside of the test tube. Other than that, it went smoothly. Dr. Dowling is very good at explaining what he wants me to do, and is very clear with his instructions. The only thing that I did not like was the pressure I felt due to the fact that he was watching what I was doing. Of course, he was doing this to assure that I knew what I was doing and that I was doing it correctly, but it still made me a bit nervous.

I'm going to the lab today, also.

Research

Tomorrow I will begin my research under Dr. Dowling. He works in the entymology department, and primarily his work is focused on sequencing the DNA of many different types of beetles in the Ozarks area of the U.S. He will show me how to extract the DNA from the beetle, and from there he will show me what exactly they do afterwards. I'm doing this research as a volunteer, so my hours are flexible. I will probably come in only a few times a week for a couple hours each time. This type of lab work requires a lot of waiting, so I will come and go as I please.

I'm excited to begin my research with him. He seems to be an easy going type of person and is very happy to have me help in his lab. I hope I like the type of work I will be doing. But, he assured me that if I become uninterested in the labwork, that he will be able to transfer me to someone else's lab that I would be more interested in.