As it is coming close to the time of my presentation. I need to think about what my presentation is going to be composed of. These past couple of weeks, I have learned so much information that I will have no chance of including them into the presentation. So I am deciding to focus on the most important things that I have learned and their purposes. I am going to be talking about the three different types of experiments that I have been doing, In Vitro Transcription(IVT), Cell-Free Translation(CFT), and BCA Assays. IVT and CFT are both procedures that will be easy to explain because they are complimentary of each other. As I have explained the two processes before. CFT is the processes of making proteins by using mRNAs as the blueprint for the construction. However in order to proceed with the CFT, we need to have mRNA to start with. This means that we would need to synthesize it. That is where IVT comes into play, IVT is the process of taking a linearized plasmid that contains the DNA sequence that we want to make the complimentary mRNA of and making a transcription reaction. The transcription reaction will then result in mRNA that is complementary to our plasmid. However, the result of the reaction is not just pure mRNA. In the sample we will also have the plasmids that we used as the template for transcription. So we then have to treat the sample to enzymes that break down the DNA and the clean the sample to get rid of the DNA and end up with mRNA with exceptional purity. After the treatment and purification is done, we proceed to use that mRNA in CFT. The CFT reaction isn't like the IVT reaction in which we need to treat it with DNAse and purify it. We only need to prep the reaction and incubate it at 37C( Body Temp.) to initiate translation. After the incubation is done and the reaction is complete, we freeze the sample until analytics will be ran on them. I personally never have observed or done this step before. The biologist that will analyize the sample will set up a western blot. I western blot is a gel electropheresis experiment in which which we place the samples in the wells of the gel and have it travel through the gel. The equipment that is used for this experiment creates a electrical field that the gel sits in and at one side the gel is positive and at the other it is negative. You would place the samples on the negatively charged portion of the field and have it travel through the gel towards the positive side. What I just explained right now is going to be the general pieces of information that I will present in my presentation. I am still figuring out how I will incorporate BCA assays into my presentation however.
To start off with this post, I would like to say that these past two weeks have been the best learning experience I have ever faced. I see myself already molding my future in a direction that will expose me more to this field. Going into this internship, I looked at this career as something I can do as a back up just in case if I didn't want to go to medical school however that idea is changing slightly. Reflecting on my goals now, the biotech industry is becoming a much larger portion of interest for me. It reshaped the path I've paved for myself to something incredibly different. I now am considering working outside of high school in a company much like the one I am at right now while I attend school. I consider a learning environment like this as something that I have to push for in my future. I have always though that the best way to learn is going to a highly credited college that teaches well however I am pushing away from that idea now. I now believe that the best learning opportunity is not at the highest credited university that forces you to take great notes but rather at a place that lets you take notes and apply them at the same time. This internship allows me to take all the notes that I've been taking and apply them to actual experiments. For my future now, I am not just looking at what school is most credited but rather what school will allow me to actually apply all the notes I've taken and knowledge I've gained and apply them to practice.
For my internship project, I am going to create a powerpoint presentation on a In Vitro Transcription experiment that I will create in the future. I feel that this will be exemplary of me because just as I stated in my last post, majority of my contributions to the company will be performing In Vitro Transcription.
A aspect of the presentation will be the intention to the experiment which will be about what the purpose, goal, and expectation of this experiment was. It will also include the protocol that I went through as well as the results. My results will be supported by graphs mostly, as well as tables. Raw statistics is generally the best way to present data however it has to be molded into a presentable form. This means that the components of the table have to be clear and the intention of the graphs have to be easily visible. To keep the progression that is happening in the company private, I will not state any of the names for any samples nor will I state the generally goal of this project. Today I had my first teacher visit at Arcturus. I had communicated, planned, and reserved time before hand with my mentor to have this meeting. The meeting went as expected. We discussed the types of work that I have done the past week and what I can look forward to doing in the future. My mentor stressed that the biggest contribution that I will make to the company is performing In Vitro Transcription. He stated that eventually, I will be independently be having tasks to complete after my training.
In Vitro Transcription is a procedure that results in mRNA as it's final product. "In Vitro" means within a test tube, meaning that we will transcribe mRNA within a test tube. The process of In Vitro Transcription starts with a template of DNA that we will eventually then mix with our other contents. Polymerase also will be in the mix. The polymerase are what transcribes and pieces together the mRNA. They attach to the promoter site on the DNA template then travels along the template creating the mRNA simultaneously. Some other points that my mentor mentioned in our meeting was the analysis of an experiment. He stressed that a experiment will not be considered complete until the analysis and write up is complete for it. Luckily, I have already practiced creating a analysis for a experiment I've done earlier this week, I will be able to use that analysis as a reference for my future write-ups that I will make. He also mentioned that I will work with In Vitro Translation. In Vitro Translation, just like In Vitro Transcription, will be performed within a sample tube however instead of producing the mRNAs, we will mix them with lysate to result in the production of proteins from our mRNA. My overall thoughts on the meeting was that it went well. Both my teacher and mentor seemed like they were on the same page of what is expected of this internship and of me. I am excited to see what the future has in store for me. My first day at Arcturus was both astonishing and unexpectedly slow. I was a bit nervous to see how it went because my mentor wasn't going to be there. He informed me two weeks ago that he was going to be out of town for the first week of my program and that he would have liked me to start a week later for when he came back. Unfortunately that was not acceptable by school so my mentor and I organized a plan for me for the first week. I walked in today knowing that I will most likely not be seeing much lab time this week.
When I first arrived at Arcturus, I was greeted my Mrs. Perkins. Mrs. Perkins is the Executive Assistant at Arcturus, she walked me around the office and introduced me to some of the workers in the company. They were all incredibly friendly and welcoming. Mrs. Perkins then showed me my cubical. This was were I would be doing majority of my work outside of the lab. It was next to hers coincidentally. She then told me about the accommodations in the office that I can use, as well as how to use the card key that I received. It was exciting to hear that I will have access to all around the company. Unfortunately after I settled in, there was confusion in understanding what I was going to be doing today. The executive assistant didn't know what I had to do. Non of the members of my mentor's team were sure about my agenda for the day. As a result, I spent the day at my cubical studying the procedures that I was going to be ask to perform. It was a bit of a tedious task but I am happy to be able to expect more from starting next week. Having the day to study also was not terrible. The topics that I have been asked to familiarize myself with are complex topics that require a lot of time to absorb and comprehend completely. Even though I spent quite a bit of time familiarizing myself with the topics, the extra time I had today was still beneficial. I am curious to see what the rest of the week will be composed of for me. For my internship, I am excited to say that I will be going to have the opportunity to work at Arcturus Therapeutics. Arcturus is a biotechnology company that focuses its pharmaceutical advancements around the development of treatments for genetic based diseases. My mentor is going to be the director of drug discovery and I will be working on a team under his management. I will also be having a supervisor that will be looking over my work and will be my main source of assistance for when I need help in completing a lab. For my meeting that I had with Dr. Cobaugh last week, he informed me that I will be taking part in the tests and experiments that his team will be doing. Priorities will change throughout the weeks however one thing that he stressed was that I will have my own set of responsibilities and expectations. Majority of the tasks that I will be completing will be including genetic analysing and PCR(polymerase chain reactions). After my meeting with Dr. Cobaugh, I became excited for many things. I loved the idea that I will be placed in a working situation with expectations that can be relatable to my colleagues around me. It was exciting for me to hear that I will also be receiving my own bench to work on and partial responsibility in ordering the supplies and reagents that are going to be used in the experiments. However the most intriguing part about this opportunity is the fact that I will have a better understanding in the life of a researcher. Considering biological research as a potential career in the future, I want to receive a first hand experience in how things operate. Just by my meeting with Dr. Cobaugh, I already have learned a lot about the career path. At this very moment right now, I am filled with excitement and eagerness to start internship. Luckily, I don’t feel nervous or worried about not providing and meeting the expectation that have been set in place for me. I have been preparing for this experience for the past couple of months and I am ready to put as much effort needed to make this experience as successful and productive as possible for both me and my company.
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