After receiving the invitation from the team WORLDSHAPER-HZ, we were quite happy and accepted this opportunity to attend the 2022 iGEM meetup at August 7th hosted in Hangzhou. But we decided to attend the meetup online due to the conflicting arrangements that we were located in different cities.
The team collaboration invited iGEM teams from high school track and college track around the world. Through this massive online communication, we feel the scientific collaborative cohesiveness of the college teams in terms of their project schedules. In particular, we fully realize that the college students participating in iGEM have shown strong abilities in social vision, professional knowledge, and comprehensive skills, and their business plans have been very close to a mature and complete standardized business plan. At the same time, other high school students also presented to us their extremely rich creativity and imagination of Promotion Videos, some of them turned into very rich professional papers made into utility-rich science videos, and some directors plotted fascinating topics in a clear theater which brought us a lot of inspiration. In addition, our peripheral goods design and charity sale practice have also been recognized by other teams.

Our team members launched online collaboration meetup with the Lip-oceil team (SubCat_China) from Beijing on August 6th, 2022. In this online meeting, the teams of both sides introduced the team project and the completion of their current tasks to each other and jointly solved some problems in their work during the discussion.

In this online meeting, the teams of both sides introduced the completion of their current tasks to each other and jointly solved some problems in their work during the discussion. At the beginning of the meeting, the two teams briefly explained their research topics and discussed product promotion, public education, and online promotion strategies. During the process, our team also raised questions to the Lip-oceil team, such as: in order to promote team progress, how can dry lab and wet lab communicate more efficiently? What content can be included in online publicity so as to raise public awareness of health issues? The Lip-oceil team members discussed these questions with us patiently. Their answers are helpful for us to improve our subsequent publicity and the cooperation of the dry and wet labs. In response to their questions about the promotion video, our PV director explained the inspiration and shooting skills. Team Lip-oceil showed their appreciation and in reture they also shared their idea of promotion video shooting then we also provided our suggestion on conceptualizing the main theme of their Promotion Video.

Plan β (PINGHE) is a team dedicated to synthesizing a large amount of medium-chain fatty acids in vitro using the fatty acid β-oxidation pathway. Since we all use the a4milGFP gene for detection, we have many points of communication in experimental operation.

Plan β (PINGHE) is a team dedicated to synthesizing a large amount of medium-chain fatty acids in vitro using the fatty acid β-oxidation pathway. Since we all use the a4milGFP gene for detection, we have many points of communication in experimental operation.
We have had a long discussion with the Plan β team on the experimental operation. After both introducing the main expected results of our project and the general experimental process, the two teams discussed problems and solutions we encountered in experiments for a long time. Meanwhile, both teams gained some inspiration.
1. Proper preservation of biological reagents. Because we did not store the ampicillin antibiotic solution at minus 20 degrees, the ampicillin resistance was weakened, so after plating on a solid LB medium, instead of a single clone, a piece of bacteria grew, which eventually led to the failure of the experiment. It gave us some inspiration. We realized that some biological reagents, such as antibiotics, host bacterial cells,s, and enzymes, would be inactivated or denatured at room temperature or after repeated freezing and thawing. Our follow-up experiments focused on the proper preservation of biological reagents.
2. Analytical methods in the face of experimental error. After plasmid transformation and successful monoclonal growth, we were faced with the fact that identification PCR was inconclusive. We list all elements in the identification PCR system, including bacterial broth, primers, double distilled water, and taq enzyme. We first ruled out the primers’ problem according to the characteristics of the biological parts, and then redo the identification PCR by replacing the taq enzyme with the prime start enzyme, but there is still no result to rule out the problem of the enzyme. The problem of double distilled water is ruled out by measuring the pH value,so the problem is locked. The steps were analyzed back after the bacterial solution, and a large number of blanks in the PCR identification results ruled out the problem of empty ligation of T4 ligase. Finally, it was determined that the most likely mistake was that the fragment was not cut completely during the one-step enzyme cleavage before the plasmid ligation. Our approach to analysis based on the properties of biological parts and existing experimental data inspired them. When the other team encountered the problem that the nucleic acid gel ran without results in the experiment, they used the same method to analyze the error as not correctly querying the extension rate of the DNA polymerase, resulting in a too short PCR extension time. After re-PCR, the gel was successfully run, and on this basis, the opposite side also successfully taught us how to query the experimental protocol on the official website according to the company name and reagent name.
3. We have to deal with the dialectical relationship between theoretical values and empirical values. After IPTG induction of glutamic acid, first, when we shook, we changed the theoretical value from 18 degrees Celsius to 25 degrees Celsius because there was no suitable temperature. Secondly, due to time constraints, we took it out, then measured the enzyme activity, and carried out protein purification, when the shaking time was less than 12 hours. In the end, there is only a marker on the protein gel, and there is no target protein from the first supernatant, hence the experiment is forced to restart. This experiment inspired both teams when doing experiments, not to rely too much on experience values and to operate within a reasonable range, otherwise, there is a high probability of failure. On this basis, when we neglected the step of adding isopropanol in the gel recovery, the Pinghe team gave a suggestion to add isopropanol and increase the centrifugation temperature to improve the yield according to the official protocol of the kit. In the end, our gel recovery was successful.
Through long-term communications, the two teams not only gained a dialectical view of theoretical and empirical values, further standardization of experimental operations, and a further understanding of theoretical knowledge, but also saw each other's persistence in experiments after numerous overhauls, and gained a positive attitude in the face of scientific failure in the future. Experiment failure is inevitable due to mistakes and established probability, but we should still keep enthusiasm in rational analysis, trial and error.