Team:NYU Abu Dhabi
The most common type of bacterial infection stems from contact with Escherichia coli, which when ingested can cause a variety of symptoms ranging from nausea to diarrhea. Shiga toxin-producing E. coli (STECs) are responsible for the majority of foodborne E. coli infections because the shiga toxin produced inhibits protein synthesis in all cells. While most countries now have stringent food safety regulations in place to prevent the sale of contaminated foods, small scale manufacturers, particularly street food vendors, often do not have access, time or pressure to consult laboratories about the safety of their food. Therefore, STEC-illnesses are still a major problem in countries that revolve around street food.
Our project aims to produce a portable device that allows for the detection of STEC through the use of loop-mediated isothermal amplification (LAMP), a technique that is similar to, but more sensitive than, polymerase chain reaction (PCR). The end goal of our project is to provide food vendors an opportunity to easily and quickly detect for the presence of STEC in their products to ensure that they are complying with government standards efficiently and conveniently. The results of each test will be uploaded into a database that provides consumers with the date, location and result of each STEC test. This will ensure that both vendor and consumer are safe, leading to a decreased incidence of foodborne E.coli infections.
About Us
Meet the team
Adrienne Chang
Major: Chemistry (Biochemistry)
Year: 2018
Where are you from: US/Taiwan
Something interesting about yourself: My spirit animal is a sloth.
Why do you want to participate in iGEM? I’ve always been interested in engineering and was curious to see how I could combine both passions.
Alex Cho
Major: Biology
Year: 2020
Where are you from: South Korea
Something interesting about yourself: I’m a South Korean soldier next year.
Why do you want to participate in iGEM? You don’t say no to these things.
Diego Kleiman
Major: Biology
Year: 2020
Where are you from: Argentina
Something interesting about yourself: I am interested in exploring what can be learnt at the interface between biology and other disciplines.
Why do you want to participate in iGEM? I think that iGEM is a great opportunity to develop an interdisciplinary project that draws upon biological knowledge to solve a real-world problem.
Fathurur Said
Major: Electrical Engineering
Year: 2019
Where are you from: Indonesia
Something interesting about yourself: I was 5 days away from getting struck by Tsunami.
Why do you want to participate in iGEM? Who says engineers can’t join a Biology competition?
Khairunnisa Mentari Semesta
Major: Biology
Year: 2018
Where are you from: Indonesia
Something interesting about yourself: In my life I’ve survived multiple earthquakes and volcanic eruptions… #Indonesian
Why do you want to participate in iGEM? My synthetic biology class introduced me to iGEM - I couldn’t wait to get hands-on experience in my final year!
Laura Karpauskaite
Major: Biology
Year: 2020
Where are you from: Lithuania
Something interesting about yourself: Went to a 30km cross-country skiing marathon without knowing how to ski.
Why do you want to participate in iGEM? It seemed like a great opportunity to learn more about synthetic biology.
Muhammad Shehryar Hamid
Major: Civil Engineering (Biochemistry)
Year: 2020
Where are you from: Pakistan
Something interesting about yourself: I have lived at ten different houses, each for at least more than a year, throughout my life.
Why do you want to participate in iGEM? To acknowledge myself with the meeting grounds of biology and engineering, and to learn how to integrate this knowledge in designing bioengineering systems.
Pratik Maisuria
Major: Mechanical Engineering
Year: 2019
Where are you from: Suva,Fiji
Something interesting about yourself: Not many people can find Fiji on the map.
Why do you want to participate in iGEM? YI get to work with machines and organisms, experimenting and creating a hybrid of life and automata.
Saad Sultan
Major: Mechanical Engineering
Year: 2019
Where are you from: Faisalabad, Pakistan
Something interesting about yourself: Snorkeled with Sharks, sea turtles and dolphins at three different reefs in the Maldives.
Why do you want to participate in iGEM? I’ve always had a love for biology and, well, biology and engineering together just sound irresistible so I couldn’t hold myself back.
Sampanna Bhattarai
Major: Computer Engineering
Year: 2020
Where are you from: Kathmandu, Nepal
Something interesting about yourself: I haven’t seen snowfall yet (I’m from Nepal).
Why do you want to participate in iGEM? The array of experiences required for iGEM, not only confined synthetic biology, instigated me to work on this project.
Under Construction
Introduction
The Fourth International InterLaboratory Measurement Study seeks to establish a reproducible plate reader-based GFP measurement protocol. Eight plasmids were transformed into competent DH5a E.coli cells and expressed for varying lengths of time. The optical density and fluorescence of each device was recorded over a 6 hour period. The major goal of this collective effort is to answer the following question: how close can the numbers be when fluorescence is measured all around the world?
Materials and Methods
In this Interlab study, the following 8 RBS devices were provided in the Kit Plate 7 in the 2017 Distribution Kit:
- Positive control
- Negative control
- Test Device 1: J23101.BCD2.E0040.B0015
- Test Device 2: J23106.BCD2.E0040.B0015
- Test Device 3: J23117.BCD2.E0040.B0015
- Test Device 4: J23101+I13504
- Test Device 5: J23106+I13504
- Test Device 6: J23117+I13504
The transformation protocol was adapted from the iGEM protocol, which can be found here
. All of the devices were transformed and cultured in LB (Luria Bertani) media containing 25 mg/mL chloramphenicol.
The following adjustments were made to the protocol:
- 2 uL DNA were added to 50 uL competent DH5a
- The incubation times on ice before and after the heat shock were 20 minutes and 2 minutes, respectively.
- The heat shock was performed for 90 seconds instead of 50 seconds.
- 800 uL SOC broth were added to each transformation.
- Test Device 3: J23117.BCD2.E0040.B0015
- After the one-hour shaking incubation, each transformation tube was centrifuged for 1 minute, 13,000 rpm. Afterwards, 700 uL supernatant were discarded and the pellet was resuspended with the remaining 100 uL before being plated onto LB agar plate.
On the next day, two colonies were picked from each plate and inoculated into two 5 mL cultures, yielding 16 cultures in total. The cultures were subsequently incubated at 37 ºC and 220 rpm for 18 hours. Afterwards, the OD600 of the overnight cultures were measured using a spectrophotometer. The cultures were diluted to OD600 of 0.02 in 12 mL LB medium and subsequently incubated at 37 ºC and 220 rpm. At t = 0, 2, 4, and 6 hours, 500 uL sample from each culture was saved on ice, yielding 64 samples in the end. The dilution calculation can be found here.
The 64 culture samples were transferred onto a clear, flat-bottomed 96-well plate according to the layout found here.
The OD600 absorption and fluorescence spectroscopy were measured using Synergy H1 Hybrid Multi-Mode Microplate Reader. The machine was previously calibrated using LUDOX. A fluorescein standard curve was also obtained under the same settings that were used to measure the culture samples.
Results
The results of the Interlab Study are tabulated here.
