This is great :(

Well this is great, Last week I came to find out that one of the refrigerators in DB 107 broke down and with out knowing I continued to the next part in my experiment. As I finished making my PCR master mix I got up and begun to look for my DNA extraction samples that I had left in the fridge, as I opened the fridge it was completely empty which was strange, I searched for Matt to ask him if he had moved or seen my samples which then he followed by saying oh! your samples were in there? I was disappointed when he informed me that the fridge had broken down sometime last week and no one noticed until a couple days later when everything on the inside had gone to waste. Now I have to extract the same DNA again so I can proceed to running my PCR. Hopefully I am able to get it done quickly so that I may get results fast.

Protocol

PCR protocol:
PCR mix
DNA 1μl
Primer 1 2μl
Primer 2 2μl
Master mix 12μl
DNase free H2O 7μl
Total= 25μl

Cycle
1. 95°c for 3 minutes
2. 95°c for 30 seconds
3. 58°c for 40 seconds                  Repeat 2-4 35times
4. 72°c for 1 minutes
5. 72°c for 10 minutes
6. Hold at 12°c 

Well that's my PCR protocol with the cycles included but we might need to recalculate one of the temperatures because it was just and educated guess. This week we had our rough draft for our research paper due and I found it quite difficult to accomplish since I don't have any results that can help me with my conclusion but I was however able to finish part if it. Last year when I wrote my first research paper it was quite easy because I had all this information in my head and all the results it was just a matter of putting it all together, now don't get me wrong I don't think it was the best paper ever but it was a pretty good first try. Next week I will be getting some results from my project and hopefully I will be able to revise my paper and make it even better.

PCR

Currently I have 4 plants that I have extracted DNA. With those four samples I am now able to run it through the PCR to amplify the DNA the only problem is that I have no protocol for the PCR yet. The protocol is currently a work in progress as well as getting the primers ready since the primers come dry (As in they look like a powder). I know Jervana is also eager to get those things as well. As soon as I get them I will begin my PCR and hopefully yield promising results. I almost forgot that this week I am suppose to have 90 hours completed for S-STEM and I am a bit behind on that so now I will have to be at lab for a while to catch up. The picture below is how I feel right now with all the stress of classes and work.

(iFunny)

Second attempt

The extraction for the the tomato and the carrot came out really well but there was a slight problem. When I was extracting the DNA from the tomato, I didn't yield a high amount of DNA so I had roughly 0.1mL of tomato DNA and after running it through the Gel I was left with practically nothing. So the bad news is that I will have to extract more DNA from the tomato. The second part of the bad news is that when I was pippetting I accidentally tore the well in the gel where I was placing the DNA and didn't notice it so nothing came up on the gel except for the carrots DNA. Next step is to extract more DNA from more plants and then run then through the PCR to amplify it.

Good news

Well the good news is that the wizard kit actually worked and although it was a bit difficult to understand once I got it done the first time it was quite easy after that. I ran the extracted DNA in the gel box and there was some DNA present so that was pretty exciting. At first when I saw the gel it was quite disappointing but looks can be deceiving a closer look at it under a UV light showed that there was indeed some DNA present in my samples. The samples present below were that of a lettuce leaf. Now that I know it works it is time to extract more DNA from different vegetable and fruits.  

The Struggle

Well this past week hasn't been all that easy for me going back and forth on what I want to do. I have been an intern here at phoenix college for about two years now which is a very long time. Currently I have no clue if I want to continue this internship so I spoke with Amanda. I'm currently trying to find something that I'm passionate about which I have yet to find. It's a big struggle for me to find something which I am passionate about. I have done a wide range of jobs, internships, and volunteer work solely for the purpose of finding something that is interesting to me. Even though I'm in this dilemma I will strive to do my best with everything, don't get me wrong I love being in the Biosciences department but I think I may need a change of environment or maybe I'm just freaking out and there is nothing to worry about.

Electrophoresis

Well last week I began my DNA extraction protocol and found it quite easy to do, or so I thought. When the time came to using the Wizard kit it was difficult to follow the protocol that came with the kit so I was asking Matt tons of questions regarding how to use it. After a while of going back and forth with Matt I was able to write a protocol in my own words. At first it took me quite some time to get used to the process but I believe that after a while I will be able to do it in a much faster pace since everything is practically repetitive. Tomorrow I will be running my extracted DNA though a gel to find out if I have any type of DNA present in my sample. When that is complete I will be running the rest of my sample through a PCR machine.