. NoteBook .

- We are starting to select new members of XMU-China 2024! So excited to start a New Year of synthetic biology - post a photo of the introductory lecture!

- The eve of the 2023 to 2024 New Year's Eve coincided with the deadline for submitting our second selection assignment. While everyone was celebrating the New Year's Eve, a group of students who were aspiring and expecting to be selected into iGEM, namely us, were studying hard. We were so busy during this wonderful time that we even declined an invitation from our classmates to go to the playground to watch a meteor shower, what a pity! But fortunately, finally, after successfully completing it, we managed to get together with everyone in the middle of the night and spent a few hours having fun with them. But it also means that we're also coming up to the final week of the semester, and time is tight!

- Wow, we can put these DNA fragment together through the sticky ends of the enzyme cutting site, amazing! That's a real boost to our knowledge!

- At last! After such a long time of selection, the new team of XMU-China is completed. Next is to determine the project, go! ! !

- Today was our first in-person team meeting, and it was so exciting to finally meet everyone! After our discussions, we headed to the Cat Street Courtyard for some food and drinks. It was such a joy to gather together, share laughs, and enjoy each other's company. We were really looking forward to our upcoming competition and all the great moments we'll create as a team. What a fantastic start to our journey!

- It was so difficult to do our endnote homework today. Some of us totally didn't know how to insert the literature into manuscript! Through all kinds of learning and consulting, we finally completed it after fighting for several hours!

- This week, all of the members of the lab group in our team had wiki assignments. This was our first exposure to computer programming, and the process of writing code was a little bit fun but also hard to learn.

- Recently, we started to draw schematics of our project, We try to use our imagination to describe the principle vividly, such as enzyme dolls, scissor hands, grasping hands and so on.

- Today, We conducted an online interview with Prof. Liang.

- We worked together on interviewing Xiamen citizens to learn their demand for writing correction products, and it was very rewarding.

- On this day, all of us from Xiang'an Campus woke up early and headed to Siming Campus for our experimental training session. It was challenging yet incredibly enjoyable. Before we realized what was going to happen, we first heard the following words and thought them very funny. For example, "If you feel stressed, then go to the lab, because there are autoclaves that are more stressing than you"......

- In this team meeting, we reported on our whole project design. We were looking forward to the experimental days ahead to start a new chapter!

- What an atmospheric picture! Following the experimental training and the team meeting, everyone celebrated Chenxin Xu's birthday in grand style, much to her pleasant surprise. Even the online members sent their warmest wishes. It was a truly unforgettable and memorable experience that we will cherish forever. Chenxin Xu expressed her sincerest thanks to everyone and XMU-China, this big warm family.

- It's raining tonight, and we are quite exhausted. However, when thinking about the interesting human practice activities we participated in and the valuable information we gained in Zhangzhou, we feel very happy!

- It's all been so bumpy recently. Finally, at this day we completed a 17-page, 7701-word lab protocol on enzymes and deinking!

- During DNA Gel Electrophoresis experiment, one of us found that the DNA bands were not migrating during electrophoresis. After three attempts, the issue was identified as using 50x TAE stock instead of 1x TAE buffer. The experiment ended successfully after correcting the buffer concentration to 1x TAE.

- At this day, one of us made the most of her time and tried making competent cells by herself in the laboratory in the Tuoyuan Building for the first time. Fortunately, what's truly worth celebrating is that the competent cells she made over these two days were both successful!

- Yesterday when one of us extracted the plasmid from bacteria, she found that the bacterial content was horribly low, mainly because the bacteria in the Tuoyuan Building grow surprisingly slowly. Well, we'll just have to continue today. Fortunately, the final concentration we measured was ideal, saving us from repeating the experiment.

- Our strain carrying the constitutive GFP expression plasmid has grown up. It seems that cells with a single plasmid tend to grow more smoothly.

- Take a look at our group meeting routine!

- At this day, we had an exchange and learning session with HUBU-China. We not only introduced our respective projects but also shared the experiences accumulated by our teams. It was the first time that we exchanged ideas with other iGEM teams during this season. Therefore, our team took it very seriously, with everyone participating, making for a grand scene. Happily, the overall communication went smoothly.

- Everyone celebrated birthday for Mingzheng Fang and Jingwen Wang!!!!!

- Some of our trivial experimental routines - determining OD₆₀₀ values.

- The final exam week is over. Next, all team members, please dive into the team work immediately, ha ha!

- After the team meeting, we had a dinner together to relax, and everyone showed off their parrot-like singing voices without stinting. In the midst of the fun, everyone deepened their impression of each other.

- Come and take a look at our homemade series of pulp reaction devices! Isn't it very interesting? There is no doubting that they can be definitely considered our original creation!

- The third semester began! Being the first team member to shuttle between Xiang'an and Siming campuses to do experiments, it was also a challenge for ChenXin Xu. As you can see in the picture, at this point she hadn't even started working with sfGFP, an amazing fluorescent protein, and she was still conducting experiments with constitutive fluorescent protein expression vetor.

- Today's experimental phenomenon seems to be working well, and the contrast is relatively obvious.

- It were the recycled paper products we produced after working tirelessly without rest.

- Let's leave a record of our today's achievement.

- Today, one of us went to visit another lab, which is doing experiments on Pseudomonas putida. And she found that the smell of E. coli is still very good.

- Recently, we have been holding our lab group meeting per Thursday at the Siming laboratory, and our experimental schedules have become so detailed that we outline specific tasks for each day. It has to be said, the coming weeks are going to be very busy.

- Today, Jia Li and ChenXin Xu went to Huaqiao University to learn how to make 3D printed hydrogels. It was so hard to explore, but overall we felt fresh.

- After discovering a new experimental direction, this is our first time conducting a validation experiment related to an inducible sfGFP-based autolysis system. We hope that the experiment goes smoothly.

- At this day, Jia Li and Chenxin Xu stayed up all night in Lounge 494 and Lab 561 to write our lectures, which is the first time we've worked so late together, and also kicked off our team's first all-night trip.

- Let us experience the scenery — the morning light from outside our experimental base.

- BBa_K5136201, BBa_K5136202, BBa_K5136203, BBa_K5136204, BBa_K5136205 were constructed successfully.

- The CCiC journey has left me incredibly excited! We met many like-minded friends and had in-depth discussions about our topics, gaining valuable advice. This experience made us realize that iGEM is not just a competition; it's a warm and supportive family. We truly feel the passion and strength here. iGEMers, let's keep pushing forward together and make a positive change in the world with our efforts!

- After the new primers were delivered, we immediately started the experiments. The PCR results for constructing the new circuit were really great with the bright and thick bands, indicating that it's a very successful experiment! Indeed, when experiments go well, everything around us seems beautiful, even the color of the gel used for recovery looks particularly nice.

- Tonight, it's another all-nighter in the lab. Let me show you some green fluorescent proteins. They are so brilliant that they almost hurt my eyes! I'd like to temporarily call them "the brightest proteins of the season." (PS: The one on the left is GFP, and the other one on the right is sfGFP.)

- Looking at the fluorescence from a different angle, it's so cute!

- BBa_K5136224, BBa_K5136225 were constructed successfully.

- BBa_K5136226 was constructed successfully.

- Due to participating in this competition, a part of us have their own passports for the first time!

- After trying countless ways to separate the ink from the pulp, today we finally found that we can extract it with limonene. To our surprise, the effect is very good, and finally we are not heartbroken persons!

- Today is one of us first time staying overnight in the lab, and she felt a little excited! She is not sure if she'll have more nights like this in the future, but she is committed to finishing all her tasks for this season. She hope there aren't too many more late nights ahead, haha, because health is really important! The atmosphere tonight is great, and She is looking forward to the results!

- We came up with the idea of creating our promotional video in the form of a conversation between a witch and team members.

- BBa_K5136230 was constructed successfully.

- Discussing topics between our teammates until dawn, rising not only the sun but also our hope for the future!

- BBa_K5136227 was constructed successfully.

- BBa_K5136228 was constructed successfully.

- BBa_K5136229 was constructed successfully.

- Recording the basic operation of pouring plates in the lab.

- After such a long time of design, experiments and human practices, our project design is finally determined! The push that we send received a lot of people's attention. Ah, so happy!

- The chitosan adsorption experiment showed exciting result! The ink solution is much cleaner after chitosan adsorption compared to the control group that uses paper to adsorb.

- Recording of beautiful fluorescent proteins.

- Great! The transformation of so many plates was very successful, and the construction of the new circuit for our autolytic system is about to welcome the dawn of victory!

- Following our team meeting, we wandered around and ended up at the Midnight Diner. Here, we put aside our research topics and experiments, enjoying a pleasant time together. After dinner, members of our two groups (the Printing Group and the Life Group) engaged in a heated discussion. It really was a thorough and intense brainstorming session. In the end, we exchanged well wishes and cheered each other on with a hearty "Go, go, go! Let's ace those experiments". Onward and upward!

- BBa_K5136220, BBa_K5136221, BBa_K5136222 were constructed successfully.

- The experiment with biplasmid cells is still in progress. Today, we tried to see if there could be any other experimental protocols to validate the autolytic system, such as assessing it solely by measuring OD600. However, after three days of hard work, we found that this method is not feasible.

- At Siming Campus, three of us stayed up all night doing experiments, and we all felt quite exhausted. However, taking a break from the experiments, we enjoyed a lively and inspiring conversation, and we all share a strong hope for a fruitful season ahead.

- Today, Zimo Liu and Chenxin Xu used a new experimental method, and the effect is so obvious that we wonder what will happen to the results of the later measurement.

- After the last batch of experiments was done, it was time to start the next set of experiments without stopping. I have to say, the neatly arranged vials look pretty good when they are placed inside the shaker.

- The printer showed up today, and it's so cool to see it running! Now we can finally do some proper quantification of the ink. Can't wait to get started with our experiments!

- Today we had a collaboration with two other teams, where we showed each other the human practices we had carried out and exchanged experiences. It is worthwhile for us to learn from each team as they are so good at human practice!

- This is our new modified reaction unit for deinking pulp experiments.

- Such experiments have been done many times, even in many different kinds. We don't know how much longer we will continue to do this in the future before we can wrap it up successfully, but we are looking forward to it.

- They are glowing! Look how bright the superfolder green fluorescent protein is! They truly live up to their reputation as proteins with fluorescence values in the tens of thousands. There's no doubt that their brightness is enough to leave a deep impression on everyone.

- Today, after completing the cross-linking chitosan experiment, Zhengyang Mo had some leftover chitosan solution and he injected glutaraldehyde solution directly into the chitosan solution. To his amazement, the solution gradually transformed into a golden-yellow gel!

- Daily sleep has been seriously insufficient, leaving us both physically and mentally exhausted, but fortunately, the heavens have not failed us and our autolytic system experiment has received fairly ideal results. Let us show you our "Army of five hundred thousand" from the autolytic system!

- Recording the day-to-day activities in our lab: glue making, glue running, protein purification, and more.

- Feeling awesome today! CYP199A4 253S is doing a great job with deinking—we're finally seeing some real results after all this effort! Go, monooxygenase, go!

- This is a small portion of the lysate supernatant of deinking enzymes that came out of Chenxin Xu first protein purification today. The lab is in full swing with all kinds of experiments, building circuits, purifying proteins, cellular experiments and so on - it's a bustling scene.

- The much-needed filter paper for the experiment finally arrived, happy to explode!!!

- BBa_K5136023, BBa_K5136024, BBa_K5136025 were constructed successfully.

- We participated in an education activity at the Xiamen Science and Technology Museum, promoting waste paper recycling and environmental awareness. Some of us organized a quiz game for children on recycling and forest protection, while others took photos, distributed surveys, and gave presentations. Through teamwork, we raised public awareness and deepened our understanding of the importance of environmental education.

- During spare time at the education activity at the Xiamen Science and Technology Museum, we wandered around the science museum. A photo of Li Jia and Chenxin Xu was even displayed on the big screen.

- Exhibiting our protein purification heroes — the ultrasonic cell disruptor and the AKTA instrument.

- This is the first time Chenxin Xu doing it, and it made her feel truly magical. Turning knowledge into power!

- The protein we purified for deinking today was in a good concentration, and it was quite something to be happy about.

- Successfully constructed a new plasmid the day before school started. If the sequencing results come out, we can move on to the later experiments!

- Finally completed the second round of deinking enzyme screening QAQ, followed by saturation mutation, come on, 2024 XMU-China iGEMer go forward!!!

- Had a whim to show off Chenxin Xu's incomplete data to give you all a little shock haha. Every save is the result of scrapping, a handful of bitter tears, who knows what it's like.

- BBa_K5136223 was constructed successfully.

- Finally we finished the promotion video.

- BBa_K5136018 was constructed successfully.

- The night before, I made my debut upload of a webpage to the GitLab repository, only to mangle the code framework like a novice chef. The pipeline crashed spectacularly. After much toil and more cups of coffee than advisable, I finally cracked the code at the stroke of 2 AM. This reminds me of that line from Dark Souls III, "Long may the sun shine!"

- BBa_K5136003, BBa_K5136010, BBa_K5136012, BBa_K5136013, BBa_K5136016 were constructed successfully.

- Today is a very lucky day, my conversion experiment finally succeeded. Last week's conversion experiments were not very smooth, in some conversion experiments, no bacteria grew on the board; In other conversion experiments, a few bacteria grew out of the board, but the bacteria were picked from the board, the plasmid was extracted and sent for testing, and the sequencing results also told me that the gene of the protein had been mutated, and gfpfasta could not glow. After more than a week of hard transformation, the experiment finally succeeded, the luminous bacteria grew on the two boards, and the colony pcr also got the correct band.

- BBa_K5136015 was constructed successfully.

- The weather is sunny and the mood is good. I ate delicious food and petted kittens today! At this time, I have successfully completed the current phase of my work and am ready to start working on the next phase. To be honest, I always prefer to be alone to paint in peace and quiet.

- Yi Zhang has drawn eight schematic presentation diagrams on experimental design in the last two days, and she even feel like a war hero herself now.

- Before and after centrifugation - what a stark contrast.

- These days we finished the first version of the web page, but was told that it needed to be overhauled and that it was time to rewrite the code.

- Today, after the team meeting, we went to dinner together. We discussed something interesting in the experiment and enjoyed a variety of delicious dishes, such as big plate chicken, sliced fish, and garlic shrimp. The restaurant also offered an ice cream buffet, where I created a huge ice cream cone that was both sweet and delicious.

- Finally, all 20 proteins were purified! My body and mind are refreshed, and at the same time, my sense of accomplishment is up up up!

- Accidents do happen — today, Yifei Yin hurt her leg.

- BBa_K5136005 was constructed successfully.

- Today, we happened to work on protein-related experiments. It has to be said that we have become quite skilled at basic operations such as preparing protein gels and running gels.

- The weather is rainy and Yi Zhang not in the mood at all. However, after the group meeting today, we all got together to chat, which made me feel extraordinarily relaxed. Next, she is going to continue working on drawings and look forward to finishing the webpage as quickly as possible these days.

- At last! The saturation mutation of CYP199A4 is complete, and the next step is to try to combine the secretion system with the deinking enzyme. Can we really accomplish this?

Whoa, whoa, whoa! LMT-253E circuit has been built! We can finally try the effect of deinking enzyme in combination with secretion system! Unfortunately, the circuit build of LMT-253A failed, and it may not be possible to build it before wiki freeze.

- Today, we visited Yanwu Elementary School in Xiamen and had a wonderful time interacting with the kids. We taught them about the importance of protecting forests and saving paper, and they were incredibly enthusiastic and eager to learn. We even conducted some fun experiments together, and the kids were so cooperative and engaged. It was a delightful experience, and their excitement was truly contagious. We left feeling inspired and hopeful for the future, knowing that these young minds are so receptive to environmental conservation!

- That's great! Although 253E is not the best deinking enzyme, LMT-253E still performs well. Let us work harder to see LMT-253A performance. We look forward to showing its better deinking effect in the presentation video.