Team:SIS Korea/Experiment
Labnotes
Over the course of the project we have taken notes of the general time period each of our event or action was completed. This summarizes our lab notes and our human practices.
2018
October
Education about the basic premise of our project
With the help of our instructor, we started off by learning about the plastic waste crisis and how to get rid of plastic so that we could set up a rough plan for the next few months.
Brainstorming of ideas for the construction of the parts
Talked about what kind of genes we were going to target, what kind of plastics were we going to combat and what genes are compatible, etc.
November
Education about various lab procedures to come
We learned about what the procedures we were going to have to conduct in order to develop our project.
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Miniprep
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Ligation
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Gel Extraction
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Transformation
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Sequencing
Go to Incheon University for discussions relating to plastic
Discussed the characteristics of plastic, and what potential biological solutions there are to solve its consequences
December (Winter Break)
Training for the cloning process
We started cloning the various genetic parts to prepare for the intensive labs to come.
Learned and practiced other necessary procedures of molecular biology
Western Blot, SDS-PAGE, etc.
2019
January ~ April
Molecular Biology Meetings
Met up once per fortnight at school to discuss additional information of molecular biology necessary for understanding our project
Collected academic papers for expanding our comprehension
Decided on the main biological components of our project
Settled on using light induced promoters (pDawn) and constitutive promoters
Decided to use PETase and MHETase as our main method
May
Requested gene synthesis
Requested from Twist Bioscience, IDT DNA
June
Checked promoter activities for both activities
Using light inducible promoters and constitutive promoter, we constructed 12 different parts with different components
June 16
Dr. Gert Weber
Dr. Gert Weber studies mostly about molecular cloning and gene expression, which are the main subject of the biology part of our project. Dr. Weber taught us the technicalities behind how proteins can be broken apart to see the amino acid sequences to change them to increase gene expression. He also advised us to not put the E.Coli in saltwater (a.k.a. oceans) because the protein will no longer be crystallized, denigrating the enzyme. With his advice, we changed the focus of our setting from a salted lab, which we were trying and were accountering some difficulties to begin with, to a media of pH 6.6
July
More testing
Linked the PETase gene with the promoters, tested degradation
Tried adding a ribosomal binding site, and tested secretion
Used western blot, SDS-PAGE
July 29
Collaboration with Korea_HS & KOREA
While looking for possible teams to collaborate with, we were approached by Korea_HS for collaboration among all three highschool Korean teams. (We received a message in June) After deciding on a day, we met up with Korea_HS and KOREA in the local cafe in which each team took turns to present their projects and ask for suggestions. After the presentations, each team went around, advising each of the other projects. A detailed account of this meetup can be found on our Collaborations page.
Went into proper testing of PET Degradation capabilities
August 13
Dr. Aleksandra Mironczuk
After reading her research paper on plastic-degrading microorganisms in marine environments, our team was intrigued to ask her about the various factors that might affect the efficacy of our bacteria in harsh, uncontrolled environments. In the end, our team learned about the different factors that lead to the formation of biofilms, which later helped us contemplate the realistic safety of the presence of biofilms on the plastics in the ocean.
August 27
Dr. Manfred Weiss
Manfred Weiss, as an expert on x-ray crystallization of proteins, helped us understand the specifics of the biology behind
August 30
Dr. Uwe. Bornscheuer
As a colleague of Dr. Gert Weber, Dr. Bornscheuer was also a lead researcher on the research on plastic degradation by PETase and MHETase. After explaining to him about our plans for our prototype, he was quick to tell us that making this into a venture would be quite impossible due to plastic being less and less productive as plastic is recycled over and over. With his contribution, we focused on creating the prototype based on the idealistic world, rather than on the business-related reality.
August 31
Tajkia Syeed Tofa
An interview with Tajkia Syeed Tofa, a researcher at KTH Royal Institute of Technology, was able to help us further deepen our existing knowledge and discover new methods of degrading microplastic (photodegradation). Dr. Tofa told us that the best way to intrigue people to lessen plastic consumption is to educate them from a young age to instill this recycling practice as a habit. With her advice, we started planning education sessions for our school’s middle school and elementary school, along with SOT’s kindergarten.
September 10
Kindergarten
We went to SOT (School of Tomorrow) kindergarten to educate the students about plastic objects used in daily life and how they affect our environment. We prepared facts & details about plastic waste, interactive games (e.g., sort the plastic), drawing pictures, and finally, a class discussion about this plastic epidemic.
September 11
Prototype
Our team started developing our final prototype model during this time period. We had a couple of members who were used to coding and robotics who helped us greatly when it came to building the final product. This process continued for a very long time as we found a lot of things we could continuously improve on.
September 16~18
Middle School Education
After contacting the middle school, our team developed a presentation and Kahoot, an online quiz service, to present to 6th, 7th, and 8th grade. Since the specifics of our biological development process were much advanced than the science learned in the middle school curriculum, we tried our best to make the biology presentation concise and brief. Instead, we focused mainly on the environmental implications of our project, namely the necessity to reduce the use of plastics.
September 23~27
School Campaign
We held a campaign in our school’s atrium, presenting posters on our iGEM topic, as well as posters on the general background of the plastic epidemic, and a survey on the public’s perception of plastic consumption. The survey results and specifics can be seen in the school campaign section on our human practices page.
October 5
Prototype finalization, testing phase
We finished the key components of the prototype and started testing its efficiency.
October 6
iCoop Exhibit
Our team held a public campaign in coordination with iCoop, an environmental group in Korea, that is widely known for holding public campaigns and education sessions about making a change and addressing the plastic epidemic. Here, we displayed posters about our project and also educated and spread information to passing tourists to raise awareness.
October 13
Jiri National Park Campaign
We collaborated with Jiri National Park, a popular wildlife tourist attraction in South Korea. Before the campaign began, we had exchanged messages and information about what we wanted to do, and we came to a consensus of exhibiting our team’s posters and banners in several locations in Jiri National Park, including the ticket booth and the entrance. The national park allowed us to display our signs for an entire month to convey the information that we aimed to share with the public.
October 14 and 21
Elementary School Education
After contacting the elementary school principal, we got connected to the SIS 5th grade spirit club. On the first day, we presented to them facts on plastic usage and what we could do to lessen it in their daily lives. To install this reusing ideology in their minds, we held another meeting on the 21st, where we made grass heads using recycled plastic cups as their vases.
October 15
Website production
Our team had been working in google docs for a long period of time, but starting from this date all our information was slowly being moved from the docs to our website. This process was very challenging as it took a very long time to format and organize all our information.
October 15
SEM data prototype
October 16
Collaboration with KUAS_Korea
We were fortunately able to contact the only Korean collegiate team, KUAS_Korea, for advice. During our collaboration, we had a mentor-mentee relationship with the team as they had participated in iGEM multiple times. They taught us what to prepare for the jamboree itself, especially related to the presentation and judges' questions.
October 18
AFM data of prototype
Reviewed efficacy of prototype comparing SEM and AFM data