To fully engage with different groups of stakeholders during our Human Practice journey, we carefully analyzed the features and vested interests of the stakeholders and, hence, characterized four categories of events to not only enrich our project implementation but also maximize the potential impacts that different stakeholders can contribute to our project.

In this section, we validate our project by carefully analyzing the suggestions and criticisms we received, ranging from the social and environmental implications of communal discussion with stakeholders. The topic of biosafety, bioethics, and the desirability of our project are all discussed. We comprehensively analyzed the diversified feedback and implemented it into the project by refining its experiment and hardware design.

In this section, we demonstrate our project has great significance. Various interviews with stakeholders ranging from the general public to specialists with relevant expertise are conducted, allowing us to understand the defects in the present control system of Giant African Snails, the lack of awareness of the general public regarding the hazard brought by Giant African Snails, thereby demonstrating our project's great significance.

Our team conducted human practice activities to address issues regarding our experiment and hardware. Throughout our iGEM journey, we interviewed various experts, discerned the pros and cons of our the technologies we might use, the defects of ctDNA as biomarkers, and subsequently formulated solutions and hardware. We also conducted evaluations on the use of science in our project to ensure the maximization of the advantages of synthetic biology.

In this section, we demonstrated our interaction with bioethics and regulation specialists. From the interviews, we were able to refine our project to be more ethically responsible, and the interviewees also influenced us and enlightened us to disseminate the idea of bioethics as a field of study as part of our education to interested listeners. Our interaction with biosafety regulation experts lets us identify the potential biosafety hazard in our project by eliciting our kill switch design. Experts also have provided us with plentiful suggestions for our biosafety proposal.

• Impetus: When developing a templated approach for the recurring problem, it is easy to be distracted by multiple aspects of view, which we need to value quality over quantity. Thus, this section demonstrates the successive impetus that propels the continuous implementation of human practice activities.

• Two-way Communication: For our solution to be human-centered, the two-way communication method is to provide stakeholders with the values we incorporate in our project design and ensure they clearly understand the project, and hence, stakeholders are able to address their concerns and suggestions precisely.

• Analysis: The two-way communication with our stakeholders is carefully analysed. In this section, feedback from stakeholders is processed and refined into constructive guidance for project implementation, allowing us to consider what work should be prioritised to best address the stakeholders' concerns.

• Implementation: Implementation reveals our actions to evaluate and refine the project, which is fully integrated with the project designs and other parts of our project to ensure our project and activities are good, responsible and responsive to the world.

Impetus
During our online research on the context of invasive species control in Shenzhen, despite the severity of the invasion, we could hardly gather more necessary information on standardized control measures for Giant African Snails. As a result, we failed to conclude the reasons why the control measures are ineffective. Some sanitation workers, as we know, are mere entities that directly involve the treatment of Giant African Snails. Thus, we conducted a street interview with sanitary workers to obtain first-hand information and new insights on their experience in tackling Giant African Snails.

Two-way Communication
The sanitation workers we interviewed informed us that Giant African Snails often appear in streets and public parks during rainy, humid days, and chemicals are used to eliminate Giant African Snails. They informed us the chemicals (Metaldehyde in particular) are more poisonous in comparison to commonly used pesticides. Meanwhile, they didn't have specific schedules or orders to eliminate the snails regularly. When we asked the workers whether they were aware of the presence of parasites in snails, only a few workers were mindful of the existence of parasites.

Analysis
Learning from our discussions with sanitary workers, the chemicals used in snail elimination could pose a serious danger to humans and animals; in particular, metaldehyde shows non-selective herbicidal properties and molluscicidal properties and is poisonous to dogs. With regard to protecting local biodiversity, we believe that directly killing the snails using toxic chemicals cannot be accepted as the optimized solution to the invasion.

Implementation
Words from sanitation workers made us realize an alternative substance for metaldehyde is needed, so we then started literature research in an attempt to seek an environmentally friendly killing agents of Giant African Snails that might be utilized in a large-scale elimination. Meanwhile, the inconveniences during sanitation work being addressed in this interview made us think more deeply about the real challenges workers might have to face in the elimination process.

Impetus
Our in-street interviews with sanitation workers provided plentiful information. Nevertheless, little information was obtained regarding the operating procedure of the elimination of Giant African Snails. Without further a due date, we again reached out to the manager in a residential community to experience the sanitation process. From this event, we aim to better comprehend what the most crucial needs in the current killing system of Giant African Snails are, and how we can design our project to most effectively address these practical issues.

Two-way Communication
We experienced a "ritual elimination" in one of our team member's residential community after a rainy day, as snails tend to be most abundant after rain. This time, we were able to wear sterilization equipment, of which the stuffiness inside the sterilization work clothes and the heavy weight of the medicine water in the backpack (weighing around 50 kilograms) exhausted us.

When we asked about their impression on Giant African Snails, the two staff members said that Giant African Snails consume plants rapidly, which induces significant harm to the community environment and ecology. However, none of them were fully aware that snails carry parasites that might significantly threaten their health if they accidentally contact the snails and are infected by A. cantonensis during their sanitation work.

We learned that in formal snail elimination, highly toxic potions, are perilous, that is, the killing would inevitably affect other snails and insects in the community and even pet dogs. This information is in accordance with our previous in-street interview.

Analysis
We believed that too much labour work involved was one of the key factors that slow down the efficiency of the control of Giant African Snails, since wearing such a set of equipment to carry out a thorough sterilization of an entire large community is undoubtedly a vast human consumption. Reaching an agreement with the manager in the residential neighbourhood, we agreed on the conclusion that an innovative measure to attract Giant African Snails can save a considerable amount of time and shrink the area of natural environment affected by the collective treatment to the snails.

Meanwhile, the lack of awareness of the threat posed by the Giant African Snail, even by the elimination practitioners, reflects the underestimation of the risk of Giant African Snails in the public's daily life, and this lack of information can cause neglectful contact with snails and lead to parasitic infection. So, there is no time to delay disseminating information about the Giant African Snail. Furthermore, we devised the idea of a set of automatic sterilization devices specifically targeting the parasites inside the snails.

Implementation
The ATTRACT and ELIMINATE procedures showed their first phototype after our sanitation experience, and we started to explore the appropriate and effective agents to attract Giant African Snails and eliminate A. cantonensis through consistent literature research. We also initiated the design of our hardware regarding two critical objectives of our project that are responsive to the needs of the sanitation workers and residential managers: Safe and Easy to use. Additionally, promoting education to raise social awareness of the problems of Giant African Snails is also heavily valued. In response, our team decided to include Education as one of the most essential portions of our implementation, starting with educating the public about the dangerousness of Giant African Snails. (See Education) After obtaining sufficient information from the public, we consider our next step to hold discussions with experts in various fields regarding invasive species to define a holistic project.

Impetus
The development of GAStroPurifier lets us identify the ATTRACT and ELIMINATE steps using genetically modified yeast, which involves human interruption to the behaviours or activities of Giant African Snails. Thus, we conducted an interview on Bioethics with Dr. Guo, who is now the senior audit expert on the Board of Supervisors and previously the chief of BGI Genomics' institutional review board.

Two-way Communication

“Be respectful, Be beneficial, Act impartial, and Do no harm.”

Values of bioethics
---Dr. Guo Zhaozheng

As we considered Bioethics related to an intricate part of our project, we inquired Dr. Guo's understanding of Bioethics. In response, Dr. Guo stated that Bioethics is fundamentally the process of seeking agreement while shelving differences with the world. From the initiation of our project, we faced the dilemma of whether we should kill Giant African Snails. After expressing our struggles with Dr. Guo, he advised us to comprehensively analyze the implications of the Giant African Snail invasion, meaning that we should not only be in the shoes of humans, but also be in the shoes of different animals, with an in-depth understanding on the harmful impacts that were obviously and significantly triggered by the invasion of Giant African Snails.

Dr. Guo, after hearing our project design, expressed his concerns about the release of genetically modified yeast into the environment. He specifically reminded us that the release would be inpreventable, and detrimental aftermath could occur.

From his interpretation, we understood that the views and opinions of people differ from one to the other based on personal backgrounds, religion, etc., and a bioethical problem shall only be viewed and judged in the present as bioethical issues vary from time to time. Additionally, Bioethics should play an essential role in both regulating and boosting scientific development; yet, it didn't proceed as smoothly as we expected, even within the scientific communities, due to various limitations and considerations.

After the interview, Mr. Guo kindly offered us a tour around the BGI laboratory, where we saw their genetically modified illuminating plants. During the tour, our team discovered several differences regarding the biosafety rules of laboratories.

Analysis
Overall, this discussion with Mr. Guo guided our project's development. We started to evaluate our project design following his instructions, primarily focusing on the assumptions of directly killing the Giant African Snails: since the Snails have invaded our city for nearly 100 years, the imprudent elimination of the species will conversely break the existing balance in the ecological system; considering human beings are not capable of - and never succeed in - fighting against the nature by killing a specific species, killing Giant African Snails is not a theortically and bioethically practical approach in the control of invasive species.

Considering the four key values of Bioethics, our project should be respectful to not only the living species, but also to the entire ecological system, and the whole project design and implementation should be as harmless as possible; being beneficial indicates that our project should maximize the benefits to the largest reach of stakeholders, without any unjust attempts that may disobey legal and moral restrictions.

Considering the four key values of Bioethics, our project should be respectful to not only the living species, but also to the entire ecological system, and the whole project design and implementation should be as harmless as possible; being beneficial indicates that our project should maximize the benefits to the largest reach of stakeholders, without any unjust attempts that may disobey legal and moral restrictions.

Due to some religious and political limitations, Bioethics is not usually open to everybody who directly or indirectly engages in scientific development, which made the limiting role of Bioethics ultimately outweigh its promoting role. Therefore, an open platform should be established to inform and involve scientific researchers in the considerations of Bioethics before launching their projects.

Implementation
Regarding evaluating the bioethical context of killing the snails, we decided to abandon the design of directly eliminating the snails and instead shifted the focus to eliminating the source of human infection, A. cantonensis. In response to the concerns in biorelease, our team started to explore and design a kill-switch system as the SWITCH to ensure the biosafety of our project. To promote the understanding of Bioethics within the scientific entities, we created a bioethics handbook (See Education) to exemplify the considerations needed when conducting synbio projects or invasive species - related projects for the iGEM community. Furthermore, we started to explore the understanding of different communities for biosafety, inspired by the tour in the BGI lab. Based on our findings, we discovered that, currently, synthetic biology laboratories have different management rules regarding biosafety. We consider a standardized rule ought to be formulated to ensure biosafety. In response, our team started to draft a new proposal (See Education) to standardize the biosafety rules in all laboratories or facilities to minimize biosafety hazards.

(Click Here for Downloading the PDF Document)

Impetus
Previous interactions with sanitation workers and the Bioethics expert provided us with new insights regarding the disease-causing A. cantonensis in snails. Posing a great threat to human health, we considered it to be one of the contributing factors to the hazardous impact of the Giant African Snail invasion, yet there is no active treatment to the infection. As we took up further exploration, we found that the operating system of parasitic infection remained unclear among online information. Thus, we contacted parasitic control expert in Shanghai Center for Disease Control to find out how the current control application(drugs) of A. cantonensis works, and the problems with the current control application for A. cantonensis and its hosts.

Two-way Communication
The interview with the expert provided our team with essential information that aided us in recognizing the severity of the infection and its reach, in particular, the L3 larvae in the intermediate host's body are highly harmful as they can invade intact skin from simple contact. Regarding how distributed this infection is, we obtained feedback that A. cantonensis can affect both animals and humans. When asking the expert about the future prospects of eliminating this parasite within Giant African Snails, we were informed that despite how widespread this issue is, there are no "specific policies" or "specialized departments" controlling the problem. Subsequently, the expert proposed that it would be beneficial if a piece of equipment helping to detect the presence of A. cantonensis incorporated into our proposed hardware. Lastly, the expert mentioned the vitality of conducting educational activities to make people aware of the dangers of Giant African Snails.

Analysis
The feedback from this interview further drove us to define our ELIMINATE step as eliminating the nematodes inside the snails for the severity of A. cantonensis infection and the lack of active control of the parasites. Inferrably, the killing agent here should be environmentally friendly with high specificity to the parasite. By eliminating the nematodes inside the snails, not only will the snails be safer for collective treatment, but also will the A. cantonensis infection rate be cut down because the disease transmission of parasitic infection will be inhibited before it involves human beings and other animal species.

The detection method of A. cantonensis, which the expert agreed with us that its application would potentially increase the efficiency of the regular screening of Giant African Snails conducted by CDC, was expected to be real-time, cheap and easy-to-operate, with biomarkers that are easy to obtain from the snails. This hypothesis reminded us of our previous project, CarcinSentry, and examined the validity of utilizing the SHERLOCK technology for parasite detection.

Implementation
To address the needs of ELIMINATE stage, we started to research the appropriate killing agents that can effectively target A. cantonensis. We also refined the previous design on cancer detection into the applications of real-time nematode detection, prompting the initiation of REASSURE stage, followed by the research of relevant biomarkers, especially cfDNA (cell-free DNA) of A. cantonensis for REASSURE detection.

Impetus
Currently, we are focused on the research of methods to satisfy our REASSURE step. From research, we did not find such detection products. Thus, suggested by experts in Shanghai CDC, we interviewed with Shenzhen Centers for Disease Control to explore the accuracy and portability of the test kits for A. cantonensis, and find out whether we can implement this synthetic biology into our hardware design. We aim to orientate the current difficulties in the procedures of A. cantonensis detection, and reflect on whether our project design is suitable for controlling the parasite and how our design may be improved.

Two-way Communication
The interviewee is the chief technician and group leader of pathogenic biology for the Shenzhen Center of Disease Control and Prevention. The information we obtained about the primary guideline in A. cantonensis control remains consistency with all other human practice activities: controlling the source of infection, terminating the route of transmission, and protecting those susceptible to being infected with the disease, including children, elderly, and those eating uncooked snails.

Regarding our ALERTS design, the expert showed great interest in the ATTRACT process and pointed out that Giant African Snails favor sweet odors, such as banana or pineapple. Besides, our group was particularly engaged with the steps and accuracy of nematode detection amongst suspected cases. The expert informed us that a highly accurate detection method is to detect pathogens on a given sample, usually tested under a microscope. Alternative methods include scanning pathological sections of the A. cantonensis-infected brain, detecting the presence of antibodies and antigens from a blood sample, and testing DNA using PCRs. Despite there being test kits available for human blood samples to be tested for the presence of the pathogen, it is not widely used in the lab, nor is it distributed much for the use of consumers. The expert questioned how our team would reassure us that A. cantonensis within the snail was successfully eliminated. We proposed the use of its cfDNA. However, the expert informed us the nucleic acid of A. cantonensis will persist in the snail body even if it is killed, causing false-positive results.

After the interview, our group was kindly offered a tour around the lab. We were shown a video of a sample of an infected cell with nematodes the lab has previously conducted. Moreover, we understood the equipment the lab uses to conduct their experiments and how each procedure was carried out in all seriousness and absolute precision.

Analysis
The expert evaluated our design to be advanced and engaging. Nevertheless, she was able to comment on the feasibility of the design. Her encouragement was inspiring. In the main, our group reached a mutual consent that the pathway and guideline for preventing the spread of the disease in the center were within our expectations, and we further understood the importance of educating those about the dangers of contact with the A. cantonensis. The expert's comment on our experimental design of using cfDNA to ensure our elimination process should be taken into consideration. The expert suggested we find a type of enzyme that would allow the examination of the presence of nematodes to be accurate. Although we were intrigued by the suggestion, it required further research.

Implementation
After the interview, we continued searching for suitable biomarkers for REASSURE procedure, mainly focused on nucleic acids and bioacitve enzymes. As the necessity and enthusiasm of ATTRACT procedure was underscored, we would like to pay more attention to the attraction of Giant African Snails by searching more about the chemical compounds with favorable aromas and their synthetic pathways. Using the information that snails prefer sweet scents, our team narrowed down our research area to substances that express sweet odor, which then turned out to be isoamyl acetate, the main fragrance of banana odor.

Impetus
So far, we have seen considerable progress on the ALERTS (ATTRACT - LOCK - ELIMINATE - REASSURE - TRANSFER - SWITCH) procedure. Therefore, we were urgent to interact with our high power-high interest stakeholders, experts in invasive species control and prevention, to gain a comprehensive review of the project design before proceeding with further implementation. Mr. Guo, the chief of the Wildlife Protection and Control Bureau of Shenzhen, who has profound knowledge of the prevention and control of invasive species and is a conservationist himself, is enthusiastic about innovative methods to prevent invasive species. From this meeting, we aim to explore the current operating system in preventing and controlling invasive species on a governmental level and obtain suggestions on our project design.

Two-way Communication
Mr. Guo informed us that Giant African Snails were commonly collected by workers using clamps and duffle bags and then sent for elimination. This method is not efficient and requires lots of labor work. He believes our project is useful for simplifying the sanitation process for sanitation workers. At the same time, he expressed his concerns on whether the presence of yeasts will block the scents and whether indigenous snails or other species will be attracted by our synthetic odors, which the necessity of selecting the snails before further operations were highlighted. He suggested the overall design can be realized more effectively if the hardware is designed, and encouraged us to conduct field research to observe the habitats of Giant African Snails and how their occurrence may affect the behaviors of other species.

Besides, what worried us was the method of disposing of the Giant African snails in hardware, which Mr. Guo said that we should not be concerned about as the sanitation workers would be responsible for collection.

Analysis
From Mr. Guo's feedback, we concluded that the current elimination of Giant African Snails is time-consuming and labor-intensive. Furthermore, we consider our hardware must be able to distinguish target species and indigenous species. Also, the hardware should ensure the scent from our genetically modified yeast spreads to a reasonable range and lasts for a relatively long period.

Implementation
First, we carried out an observation experiment of attracting Giant African Snails with homemade dough containing fresh yeasts, of which the check-in was approved by the iGEM Safety and Security Committee. For our action plan, we provided feedback from Mr. Guo to members of our hardware team. In response, the design of a separate compartment was constructed(See Hardware). Given that there was regular sanitation work related to Giant African Snails, concluded from the interview with sanitation workers and the experience of the sanitation process, we decided to make the originally automatic TRANSFER process into manual operation to ensure the effective collection of the Snails from the hardware.

Impetus
Previously, our interview with Dr. Guo prompted our understanding of the importance of biosafety. Team members responsible for education researched the biosafety regulation in other laboratories and discovered inconsistencies. We discerned a high safety hazard. Therefore, a regulation aiming to standardize the biosafety regulation in all synthetic biology laboratories locally was drafted, and we strive to educate the scientific community via this regulation. Thus, the human practice group contacted Mr. Chen, a member of the Chinese People's Political Consultative Conference. With him, we aim to discuss the proposal and know whether the proposal has high feasibility and shortcomings.

(Click Here for Downloading the PDF Document)

Two-way Communication
Mr. Chen himself, after the brief introduction of our project, claimed that he did not know snails could carry parasites. Thus, he suggested we should popularize the knowledge of the seriousness of the parasite in the snails. Mr. Chen mentioned that our project indeed has high feasibility, however, we need to inform the public that the Giant African Snails could jeopardize human health. We told him about our mini-program, and Mr. Chen, with great enthusiasm, claimed that he would help to promote the mini-program. He recommended we visit more laboratories and understand their shortcomings of the management so that we could then include the results after the visits to refine our proposal to be more persuasive and convincing.

Analysis
Mr. Chen strongly agreed that our project itself was valuable and meaningful. Our mini-program, ideally, will generate sufficient information and if we can inform and deploy a user network. This requires public education with the pamphlet produced by our education team while promoting our mini-program.

Implementation
The interview with Mr. Chen let us understand how the proposal can be implemented and the aspects we ought to consider before implementation. In response, we decided to visit more laboratories and understand their management shortcomings so that we could include the results to be more persuasive and convincing. However, the terms in the proposal still need to be revised. Therefore, we continue to contact experts who can help to revise our proposal.

Impetus
Based on our practical involvement in synthetic biology laboratories and extensive study conducted on the internet, it has come to our attention that numerous inadequacies exist in the realm of management and regulatory practices. Consequently, a collective decision was made to compose and present a proposal concerning the enhancement of synthetic biology laboratory management. After our interview with Mr. Chen, we visited the Research Center of Peking University Hospital. We interviewed Dr. Zheng to discern if there are areas for improvement regarding safety management in the biology lab, whether our proposal has a social contribution, obtain advice on our proposal writing, and information about how we can improve.

Two-way Communication
We first introduced our project and presented the reasons that motivated us to draft the proposal. He concluded the management and regulations for the synthetic biology laboratory have already been comprehensive and complete. For our reference, he also provided us with access methods with professional channels or websites, such as NMPA and NHS.

We proceeded to the laboratory located within the research center of the hospital. The introductory information elucidates that several departments within the hospital own individual laboratory facilities and employ personnel with specific expertise. Instruments are graded, and different professionals use different specialized instruments. Also, there is a designated office space on every floor specifically allocated for administration and monitoring.

Dr. Zheng told us that from the hospital research level to the enterprise level, matters large and small are regulated in detail, but the access procedures are extremely complicated, mainly due to the regulations of different departments. And that's the main reason why we cannot find related rules and regulations on Internet, and neither can those emerging enterprise-level biology labs.

Analysis
Dr. Zheng gave us some positive argument on our project, but he pointed out the irrationality of our proposal's design and misdirection. We acknowledged the current rules and regulations are comprehensive and were carefully revised. However, we discerned one drawback contributing to why we couldn’t find those rules because we didn’t get access to these platforms and channels. This raised our team's concern, as other syn-bio laboratories might also not be able to find out such information.

Implementation
Our analysis concludes that the factor that misleads us is the reason for diverse management rules in few syn-bio laboratories. Indeed, it is significantly complex to obtain authentic information regarding a regulated and compiled criterion of syn-bio management. Consequently, we altered the focus of our proposal to "Developing a platform that facilitates access to regulations pertaining to synthetic biology." This platform aims to enable individuals interested in synthetic biology or the establishment of a synthetic biology laboratory to efficiently and comprehensively obtain relevant information, helping to address the problem in the difference of biolab management, thus alleviating the potential risks that could give rise to adverse aftermaths.

(Click Here for Downloading the PDF Document)

Impetus
Our previous interview with Mr. Guo from the Wildlife Protection and Control Bureau greatly motivated the team to explore the ATTRACT procedure and design our hardware. After weeks of exploring and hardworking, the team successfully gained uplifting results in synthesizing attracting agents and designed the hardware, incorporating the needs of users referencing the response of different stakeholders in previous human practice activities. We aim to evaluate our results in ATTRACT branches and the overall hardware design.

Two-way Communication
Mr. Guo inquired us about our project progress, specifically on the attracting agents we synthesized, alpha-pinene or isoamyl acetate. We informed him that our progress of isoamyl acetate is relatively decent, though we had not synthesized alpha-pinene yet. Mr. Guo commented that it would be excited to see the substance being useful for the implementation of our project.

For hardware, Mr. Guo expressed few concerns and suggestions as we demonstrated our design blueprint. Mr. Guo asserted the hardware should include as many entrances as possible for the attraction of abundant snails. At the same time, the hardware should also be concise yet simple. He questioned our proposed design of using a blower as he believed using wind is more than suitable. Mr. Guo also stated that our implementation of hardware is most suitable in residential communities and in parks, instead of rural areas.

For our design of Wechat mini-program, Mr. Guo suggested we should incorporate the auto-collection of information and users should be able to upload pictures and locations when they spotted Giant African Snails. Also, we should educate the people if we aim to propose a mini-program to the public. To do so, Mr. Guo suggested we discuss with the Shenzhen Health Commission.

Analysis
From Mr. Guo's suggestion, we consider it is crucial to work on the simplification of our hardware. We believe users would prefer a product that is both cost-effective and efficient. The idea of removing the blower was carefully evaluated by team members. Originally, we thought the use of a blower could greatly disseminate the scent, attracting more snails. However, we soon became aware that the electricity used to power the blower may not last long. Also, we could not use solar panels as our hardware is placed in dark, humid environments.

Implementation
For implementation, we revised our hardware design according to Mr. Guo's suggestions. Additionally, a water proof test is conducted on one stormy night. Furthermore, we updated our mini-program to allow users to be able to upload pictures, and designed a background data collection method. Thus, the data we gathered via our mini-program can be used as reference for not only the implementation of hardware, but also the integrated control of Giant African Snails with a higher level of efficiency and a wider outreach.

Impetus
Investigation of the toxicity of Cry1518-35 to nematode C.elegans is essential for evaluating and verifying the bioactivity of the killing agent we synthesized for our target nematode, A. cantonensis. As we were drafting the experiment design, we suffered from great confusion and uncertainty that relevant literature was not sufficient enough to answer. Therefore, we contacted Dr. Hu from the Institute of Neuroscience of Chinese Academy of Sciences to learn about the skills in nurturing C.elegans, understand the keys to designing nematode experiments and improve our experiments as much as possible.

Two-way Communication
This discussion was conducted through emails. We first inquired about the details of nurturing C.elegans in the laboratory, and her advice helped us figure out what we might have done wrong to fail the cultivation of C.elegans. Regarding the first draft of our experiment design, she pointed out several points that could be revised or simplified, such as the number of nematodes used for each experimental group, the decision of variables and the conditions for cultivation. Overall, she also gathered her knowledge of similar toxic proteins that can potentially be utilized to enrich our project.

Analysis
Following Dr. Hu's instruction, we re-examined the details in our design with hot discussions within our team. As the ELIMINATE procedure will be conducted within Giant African Snails, we have to take the actual conditions inside the snails into serious consideration, including the growing temperature and the stage of C.elegans used for verification experiments. Meanwhile, the fact that N2 C.elegans is fed by E.coli strain OP50 enables us to conduct the experiment by the co-culture of OP50 and genetically engineered E.coli that can express Cry1518-35 proteins, which is a far more efficient approaches compared to directly testing the protein's toxicity with its concentrated supernatant.

Implementation
During and after several rounds of email interactions, we kept pace with substantial research on the effective approaches to conduct toxicity assay of cry proteins. Then, we came up with the final version of the verification experiment design and selected the optimal design to initiate the attempts in testing our synthetic protein.