202204-05
High school students from several international schools in Shanghai and Suzhou formed their own teams to prepare for the iGEM Bioengineering Competition.
202205-06
We finally started our project! Our team was divided into several groups, each exploring and proposing a possible project proposal, including a wound repair project using HA, a lactase project focusing on lactose intolerant people, and a project using bioplastic foam to make biodegradable running shoes and sneaker midsole technology. Finally, we decided that the final project is to use bioengineering to treat lactose intolerant people, considering the experimental period and target audience.
202206-07
After we decided on the topic, we started to design the experiment and use SnapGene to design the plasmid. It was also necessary to contact laboratories, purchase experimental reagents and other series of preparations, separating group members into dry and wet labs to solve problems specifically.
20220701-0714
Our team started the lab work in the first two weeks, and we encountered many problems after settling into the lab, in which the first two weeks were slow and most of the work was troubleshooting problems. We tried to extract the plasmids numbered HA0001 and HA0002 corresponding to the reagents and to draw a line culture by picking monoclonal clones of the colonies on the Petri dishes. However, due to the unfamiliarity with the new lab and the low quality of some kits, we encountered big problems in extracting plasmids: every time we ran the results of the agarose gel electrophoresis test after extracting plasmids, the results were not satisfactory. However, it is good that the experimenters summed up the lessons learned, replaced the kits in time, and ensured the standardized sterile operation of the experiment, and the success rate of the experiment slowly improved.
20220715-0731
More and more team members came to the lab, and the experiment went smoothly. After changing the kits and standardizing the operation, the success rate of our experiments gradually increased, and the extraction of plasmids, which was once considered very difficult, became easy. We also conducted more experimental follow-ups, mainly numbering lacz (lactase) and GFP (as positive control) as HA0003 and HA0004 and started the culture of lacz and GFP. Meanwhile, in late July, we started to transform the cultured monoclonal into Nissle1917 as well as DH5α, and the experimental efficiency is getting faster and faster. The stem group is also interviewing the managers of dairy companies very carefully to listen to the professionals and perfect human practice.
20220801-0814
The lab is at its most efficient and has the most things going on during this time. Experimenters are becoming more and more familiar with the lab equipment, and one experimenter can complete a day's work, which greatly improves efficiency. During this time, we reviewed the literature for extraction of podophyllotoxin (CAP) using phenol and chloroform and then designed how to prove the presence of CAP. Our original design was to use SDS-PAGE gel electrophoresis and prove the existence of CAP after running the gel, but since we wanted to finish the experiment more, faster, better, and less, we tried to stain CAP directly using various dyes such as safranin, Alcian blue and crystal violet, but the results were not very satisfactory, so we finally went back to gel electrophoresis, so to speak, in a tortuous exploration.
Meanwhile, the dry lab also started professional experimental modeling as well as wiki design, with fast progress and high efficiency. Although they also encountered certain tricky problems in modeling, the stem group members actively reflected and asked for advice to learn, solving problems with professional means, and making progress together.
20220815-0831
Near the end of the experiment, we used the kit with an electronic spectrophotometer to determine the enzymatic activity of β-galactosidase and cleverly used Excel sheet to draw a scatter plot and calculate the enzymatic activity by the formula.