Tell us about your project, describe what moves you and why this is something important for your team.

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• A clear and concise description of your project.
• A detailed explanation of why your team chose to work on this particular project.
• References and sources to document your research.
• Use illustrations and other visual resources to explain your project.

Advice on writing your Project Description

We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be consist, accurate and unambiguous in your achievements.

First of all, we use the PCR to proliferate the plasmid provided by IGEM. PCR is a kind of mechanism that proliferate the target DNA segment beyond the capacity of the nature, the velocity of the proliferation of the PCR mechanism can be up to 100,000 generations per day, thus provide us with plenty of plasmid for the experiment. Then the module is used with the purified water, 10*buffer, dNTP, F and R primer to synthesize the target opdA enzyme’s DNA. Then the result solution was put into the PCR again, waiting for later purification by using the reagent----Gel extraction kit. When waiting for the PCR with the solution we made earlier, the medium for the coli was made by mixing the soybean powder with water, the result 1 liter of solution is then equally divided into four parts because of the limitation of the container which has the maximum capacity of 250ml. Two of them are simply made by mixing water and soybean powder, while the rest two were added agar thus created two solid medium. Because of the rigid requirement of the coli and avoid contamination, the for containers with the medium are put into an autoclave to eliminate the potential contamination in the container. The next step is preparing for the separation gel by mixing the agarose and water, result in the solution with the volume of 30ml. The solution is then put into the oven to mix the agarose and the water entirely. After taking it out of the oven and cooling the solution, the dye which make the DNA appear when exciting light was lit upon the gel is put into the solution. The solution is put into the model with a comb, in order for the gel to concrete. With the previously made separation gel, the DNA model is put in to the hole left by the comb and the model is put into another hole. The electrophoresis is then used to extract the target DNA segment in the solution. The resulting gel has many stripes and one of them contains the target DNA segment. In order to purify the DNA segment, the stripe need to be neatly cut out of the gel. Then the Gel extraction kit is used to purify the DNA by following the instructions in the Gel extraction kit.

Judges like to read your wiki and know exactly what you have achieved. This is how you should think about these sections; from the point of view of the judge evaluating you at the end of the year. First lets start with the three basic components of genetic engineering: the restriction endonuclease, phosphodiester bond and the carrier in terms of the plasmid in colibacillcus. In our experiment, we used EcoR1 and spe1 as our restriction endonuclease to cut the target DNA fragment from pet32a, the fragment on pet32a is used to replace the strong promoter, RNA thermometer and ompA from last year’s project. The substitution for those three components are T7 promoter, lac operator and finally the histidine tag. With this improvement, we are able to extract the opdA enzyme directly from the E. coli. It is agreed that the E. coli is to some extent harmful to human body, so by extracting the enzyme we can readily prevent the potential harm of the E. coli. Since the histidine tag can help extract the enzyme from E. coli. The very first procedure of the project is to cut off the target DNA sequence from the model plasmid we selected, which is pet32a. We separately used EcoR1 and Spe1 to cut the sequence. Then we used PCR to proliferate the sequence. At the same time the original sequence from last year’s plasmid is cut off from the model. T4 phosphodiester bond functions as the connecting between the DNA segment and the model so the previously extracted target sequence is able to connect with the model provided by IGEM.  The pivotal procedure of the whole experiment is to transport the plasmid into E. Coli and use E. Coli as an carrier to exponentially proliferate the plasmid. First we set the E. Coli in a low temperature refrigerator which is approximately -80℃, at that temperature makes the E. Coli be in a state of sensitivity which means its membrane is vulnerable. When it is about time to transport the plasmid into the E. Coli, we took the frozen E. Coli out of the refrigerator, it immediately rises its temperature to 0℃. When the plasmid is ready, the model E. Coli is put in 42℃ environment so because of the heat affect the volume of the whole bacterium and increase the size of the membrane thus small holes began appear on the membrane, letting the plasmid in to the E. Coli. When all the aforementioned procedures are done, the E. Coli are on ice again, making the holes on the membrane close. Finally, we put the culture dish with the E. Coli plus the plasmid in to 37℃ environment letting the E. Coli continues to grow.