Team:WLC-Milwaukee/Notebook
Project Timeline
Summarized below is the sequence of events that led to the completion of our project, from brainstorming ideas to finally submitting our parts and finishing the wiki!
Project Decisions!
February 2018
Based on Judge feedback and our desire to bring the 2017 WLC-Milwaukee project to fruition, we decided to continue our 2017 project detecting E. coli in contaminated water supplies using Lambda Phage tail proteins. We began laying out the ways we would improve our past project and complete the areas where it had fallen short and been unsuccessful in 2017.
Began Cloning parts into pTrc99a
March 2018
With the help of our advisor, we designed primers and began the frustrating process of amplifying and cloning our parts from the pETail4 plasmid (containing the Lambda Phage tail protein genes) into pTrc99a in order to control protein expression levels using the pTrc99a IPTG inducible trc-promoter (also called Ptac).
MUBRC
April 2018
A number of our team members participated in MUBRC (The Milwaukee Undergraduate Biological Research Conference) by volunteering and helping facilitate setup for this event while Jack presented a poster of our past work and hopeful project improvements in order to gain feedback from other undergraduate researchers and PIs in attendance.
Started Human Practices Investigation
June 2018
As a team we began investigating issues relating to water quality and testing, how our project may fit into the market, how our project could fulfill a need for water testing in certain areas, and could be designed to reduce waste and eliminate any environmental hazards.
Organization Outreach Day
August 2018
In August Jack and Laura presented a poster and posterboard at the Organization Outreach day, welcoming new freshman students to our campus and talking with various students about bioengineering, synbio, and our project. Through this event we made contact with a number of students who were very interested in joining iGEM and have already gotten involved with our team!
Confirmed Clones in pTrc99a
September 2018
We finally obtained our clones in the pTrc99a backbone and confirmed their presence by diagnostic digest and sequencing. This was a major step as we could now clone our parts into pSB1C3 and include the trc-promoter for IPTG induction in our parts.
Science Exploration Day
September 2018
This event was a great Outreach event and was an awesome way to interact both with almost 20 grade school aged students and their parents from the local community and provide various lab activities and opportunities to these bright young students. The students participated in hands on explorations of some of the numerous areas within STEM.
Protein Purification and Analysis
October 2018
After obtaining successful clones of our parts into pSB1C3 we were finally able to start protein purification using Nickel column purification methods and analyze our parts for successful protein production.
Bacteriophages for High Schoolers
October 2018
After analyzing substrate-HRP ratios and which methods would suit our test kit system, we began a system prototype, conjugated HRP to our proteins, and performed some testing to optimize system operation both in controlled ideal lab circumstances and in more realistic circumstances to determine the viability of our product.
Submitted Parts and finished the Wiki!
October 2018
We submitted our parts to the Registry and started working on finishing our Wiki, confirming our results, and completing our project prototype!
Young Women in Science Program
March 3rd 2018
We were able to continue our relationship with the Young Women in Science Program (YWSP) providing lab activities for 13 young women interested in STEM examining bacterial membranes, membrane permeability, and bioengineering in the real world.
Planned Outreach Events
April 2018
We began laying out and deciding on some of our larger scale Outreach events for the fall and considering additional ways we could engage and educate our surrounding community in a holistic way.
New Cloning Primers
May 2018
Due to what appeared to be inconsistencies in our gene amplification we added a single base onto some of our cloning primers and increased our annealing time which allowed for better gene amplification and aiding in our cloning efforts.
Jack Tours Cornell Lab Spaces
August 2018
Over the summer, our team lead Jack was able to attend the Cornell SILS (Summer Institute of Life Sciences) symposium and meet members of the Cornell iGEM Team. He was able to speak with some of their members, particularly their team lead Rahul and tour their lab spaces. We greatly appreciate the openness and collaborations of the Cornell iGEM Team and Jack enjoyed seeing what it would be like to have an entire lab space dedicated to iGEM research!
Spoke with Experts to Analyze our System
September 2018
In September we began communicating with experts in environmental testing and the water safety industry. Due to their feedback we modified our initial project design to make it more user friendly, and altered the intended scope of our project application after re-analyzing the testing systems in place within the large scale water management industry.
Tested HRP and Substrates based on new system goals
September 2018
After re-examining our system impact and applicability we also needed to change the substrates and ratio of HRP-Substrate we had been intending on using with our test kit to better suit the niche our kit best fills. Based on these changes we began testing new combinations and concentrations of our substrates and HRP to optimize our kit in terms of both efficiency and accuracy.
Obtained clones in pSB1C3
October 2018
After much frustration we finally succeeded in obtaining successful clones of our parts both with and without the trc-promoter (also known as Ptac) in pSB1C3!
Visited MMSD treatment plant
October 2018
As another layer to our system design investigations we visited and toured the Milwaukee Municipal Sewer District (MMSD) wastewater treatment plant to learn whether or not our project would be viable on the large scale of Municipal Sewer Effluent management. Additionally, we sought to compare how our project could be used at small scale wastewater treatment facilities versus massive facilities such as the MMSD facilities in the Milwaukee area.
System Prototyping and Continued Testing
October 2018
After analyzing substrate-HRP ratios and which methods would suit our test kit system, we began a system prototype, conjugated HRP to our proteins, and performed some testing to optimize system operation both in controlled ideal lab circumstances and in more realistic circumstances to determine the viability of our product.
