Team:AHUT China/HP/Gold

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Gold

1.Expand on your silver medal activity by demonstrating how you have integrated the investigated issues into the design and/or execution of your project.

During our talk to experts, we got many helpful suggestions about the perfection of our project as well as its potential. Furthermore, they introduced plenty of new ideas in the field of synthetic biology and how they benefited us, which raised the interest of the audience. We also arranged video conferences with UESTC-Software team and NAU Team, which made the first step for our further cooperation. Hence, our business plan collected all wonderful ideas of our team members who were ceaselessly racking our brain to promote ideas to make it more practical so that we can benefit and provide services for the public in a real sense soon, which is thrilling and meaningful.

All in all, human practice rendered lots of conveniences and inspirations for us and made us know its strengths, weaknesses, opportunities and threats, all of which will motivate us to do better and we hope all our readers of this website get a general understanding of our human practice through our detailed depiction and become interested in our project. More constructive proposals are welcomed!

2.Improve the function OR characterization of a previously existing BioBrick Part or Device (created by another team, or by your own team in in a previous year of iGEM), and enter this information in the part's page on the Registry. Please see the Registry Contribution help page for help on documenting a contribution to an existing part. This part must NOT come from your team's 2016 range of part numbers.

In the project of AHUT_China in 2014, we built the shortest path calculation model based on DNA calculation, and illustrated the principle of calculation through a planning case of navigation pathway. But, this year, We have furthered study based on the path planning problem to be solved in 2014. BBa_K1341020-Solution24578, is the shortest pathway in the project of 2014 of AHUT-China, which Node NO.5 is a necessary node to be passed in order to get to Node NO.8. However, in our project of this year, we don’t have to pass Site NO.5 while taking Site NO.2 as the start point and Site NO.8 as the terminal, which provides us with more diverse pathway choices and more conforms to the real situation. In our project of 2016, the DNA single strands of the sites and lines are connected by bridging-PCR. 12 information bits are added to the DNA single strand of the lines, which the pre-4bp sequences contain the information of identification point, the mid-4bp sequences contain the information of the specified line, and the post-4bp contain the information of the path length. In order to obtain Solution24578, it needs to construct recombinant plasmid in every possible pathway. And then the recombinant plasmid was inserted in E.coli, where the fluorescent protein gene in Node NO.5 get expressed. Thus, all the pathways passing Node NO.5 screened out. By contrast, we don’t have to go through a series of complex process like what have been depicted above in order to obtain the shortest pathway <2-6-8> in this year.

3.Demonstrate a functional proof of concept of your project. Your proof of concept must consist of a BioBrick device; a single BioBrick part cannot constitute a proof of concept. (Remember, biological materials may not be taken outside the lab.)

We made some contacts with some navigation companies in order to put our project into practice and bring the function of finding the shortest pathway through bio-computing into full play. Auto Navi map received our phone call and we introduced our project to them. The way we validated the two Bio-brick, BBa_K2044000 and BBa_K204401 corresponds to the way the map advises people to take the shortest pathway plan, which the company showed great interests in our project and encouraged us to further develop our project so that the general public can afford to use it in the real life. When our project is ready to be popularized to the general public, they would love to cooperate with us. In this process, we become familiar with the operating model of some navigation companies, which make us realize what else we should do to achieve the commercial value of our project.

4.Show your project working under real-world conditions. To achieve this criterion, you should demonstrate your whole system, or a functional proof of concept working under simulated conditions in the lab (Remember, biological materials may not be taken outside the lab.)

Bridge PCR

Put all diluted sites and lines together into the PCR tube, forming all possible DNA fragments, or the different pathways.

DNA Ligation T4

DNA ligase is added to connect sites and lines.

DNA Gel extraction In the last two steps, gel extraction is needed to remove impurities.

PCR

Carry out PCR to amplify all fragments which begin with site 2 and end up with site 8.

Gel extraction

Carry out the gel electropheresis and the gel extraction, cutting respectively the gel volume of 200 bp, 250 bp and 350 bp.

Transformation and filtering of target genes

Import the target genes into the plasmids, deliver the plasmids into E.coli and coat the competent cells on the solid medium. Then screen out of the strains that we need in the medium containing chloramphenicol. After the screened strains were amplified in liquid medium, purify the plasmids and identify them. The last step is to send into the company for sequencing.