Collaboration have always been an important element for scientific research in today’s world. During the summer, we truly felt that collaboration is essential for us to get things done, in a much better way. In addition, we were also glad to help other teams.
Mentoring BNDS_CHINA - a High School Team
Since that the iGEM competition only has 10 years’ history in China, new teams without experience keep coming up in recent years. As pioneers in their own schools, they more or less have to face obstacles not only on equipment, resources and funds, but also about all kinds of knowledge concerning with basic facts, lab operations, tricks in this competition and so on. Although it is the first time for us to attend iGEM as well, we still hope to devote our scanty effort to helping other teams.
So this time, we are quite glad to mentor the high school team from Beijing National Day School (BNDS). Apart from offering some necessary reagents they need, such as restriction enzymes, we also taught them some important methods for experiments such as plasmids extraction and competent cells transformation, as well as correcting several mistakes during operations in Clean Bench (Fig. 1, 2). In addition, we also helped analyze the results they got, and gave some suggestions on their cutting and ligation protocol due to those outcomes.
The team members from Beijing National Day School have been working very hard. Beside this competition, most of them have to undertake the heavy load of Gaokao as well. They spend all the spare time staying in laboratory and consulting us with details in experiments frequently. Hope they will do a good job in Giant Jamboree.
All in all, we all felt that collaboration is quite meaningful and useful. It encourages communication between teams which brings new teams ready-made experience and creates more chances for iGEMers to make friends and share ideas. And the activity itself gives out the message that if everyone, not matter devoted to this area or just interested in Synthetic Biology, can work together and help each other, we can make a really big deal.
Helping team BIT-China
When meeting with iGEMers from Peking, BIT-China and Tianjin University, we were surprised to find that, much similar to our project, BIT-China was also working on a kill-switch based on TA modules. Characterizing varies toxins was one of the most important tasks for both teams. We measured growth curves by a 96-well microplate spectrophotometer (BioTek Synergy H5), while BIT-China used traditional methods of taking a sample from a shake incubator for every 30 minutes or so.
So we helped them measure the growth curves of bacteria harboring their toxin gene, MazF. They chose three different RBSs, and we measured the growth curves of those three different bacteria. Using the 96-well microplate spectrophotometer, the value of OD600 was measured automatically for every ten minutes, making the plot more continuous, accurate and smooth. Setting three parallels, we showed that the measurement was quite repeatable. The graph was drawn using Prism 6.0. Fig. 3 shows that if toxin protein MazF is induced when the value of OD600 lies between 0.5-0.7, it instantaneously halts the growth of the curve, indicating strong growth arrest. Interestingly, if toxin is expressed at the beginning of the measurement, as Fig. 4 illustrates, the growth of bacteria was not immediately suppressed, which is quite different from the toxins we used. The growth patterns in the first 200 minutes are not observably different from Control. This result is in accordance with BIT-China’s data.
Getting Generous Help from Nanjing-China
During CCiC at SYSU this September, we talked about our hardware design, a miniature aeration tank from waste water treatment plants. To our surprise, we got some quite useful feedbacks from Nanjing-China. They generously shared their experience of building a similar device in their last year’s project. Such details as materials for fillers have never been considered by us before. They gave us information about the effect of different fillers, which did inspire us a lot, and freed us from fruitless trial-and-error experiments. So their suggestions saved a lot of time for us, for we can spare more time on improving the design and construct of our own device.
They also provided us with their own hardware and filler (Fig. 5, 6). This greatly reduced the amount of our work. We had some technical problems while building their aeration tank, and the instructor from Nanjing-China tutored us with great patience through mobile phone.
Unfortunately, their hardware is not compatible to our project. They used Bacillus subtilis as a chassis organism last year, which naturally forms biofilm. What is more, they also transformed a biofilm protein into the bacteria, so their engineering bacteria grew well on the plastic fillers. However, we used E. coli throughout our project that does not generate biofilm. As a result, our bacteria cannot attach to the fillers after 10 hours’ incubation in a shake incubator. Although we were not able to use the device provided by Nanjing-China to test our design in real condition, we greatly thank them for advising us on building our own hardware.
Communiacation with XMU-China
Both XMU-China and UCAS iGEM teams focusing on antibiotics this year, we made our way to Xiamen for a face to face communication with XMU team in August to exchange ideas on our projects. XMU-China also gave us plenty of suggestions on project management since we are a new team.