Difference between revisions of "Team:Tsinghua-A/Demonstrate"
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<div id="apDiv15">6.Take out 200ul cultures obtained from Procedure 5 and measure the fluorescent intensity and the amount of bacteria using flowcytometry after 15h. <br> | <div id="apDiv15">6.Take out 200ul cultures obtained from Procedure 5 and measure the fluorescent intensity and the amount of bacteria using flowcytometry after 15h. <br> | ||
<br> | <br> | ||
| − | <div id="apDiv16"><img src="https:// | + | <div id="apDiv16"><img src="https://2017.igem.org/File:Tsinghua-a-killing-form2.jpg" alt="<img src="form2.jpg" width="866" height="301" />" width="618" height="302" /> <img src="https://static.igem.org/mediawiki/2017/7/79/Tsinghua-a-killing-begger.png" width="344" height="202"> </div> |
B.Test if the killing ability is adequate <br> | B.Test if the killing ability is adequate <br> | ||
5. Construct the following 8 groups and then incubate the cultures at 37℃, shaking at 220rpm. For each group, 3 replicates are constructed, numbered from 1 to 3. Each replicate contains the bacteria with the same number. All the medium contain 100ug/ml chloramphenicol. </div> | 5. Construct the following 8 groups and then incubate the cultures at 37℃, shaking at 220rpm. For each group, 3 replicates are constructed, numbered from 1 to 3. Each replicate contains the bacteria with the same number. All the medium contain 100ug/ml chloramphenicol. </div> | ||
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1.warrior 1 kills itself but warrior doesn’t .We compare the amount of warriors in medium containing A+K with the amount of warriors in medium containing C. If the amount is apparently lower in the medium containing C, the warrior must have killed itself.We find that the amount of warrior 1 is apparently lower in C group, which means that warrior 1 kills itself. Figure1 shows the result. The ratio represents the amount of warriors in C medium divided by in A+K medium.(group4/group1,group 5/group 2) *A+K stands for ampicilin+kanamycin,C stands for chloramphenicol. </div> | 1.warrior 1 kills itself but warrior doesn’t .We compare the amount of warriors in medium containing A+K with the amount of warriors in medium containing C. If the amount is apparently lower in the medium containing C, the warrior must have killed itself.We find that the amount of warrior 1 is apparently lower in C group, which means that warrior 1 kills itself. Figure1 shows the result. The ratio represents the amount of warriors in C medium divided by in A+K medium.(group4/group1,group 5/group 2) *A+K stands for ampicilin+kanamycin,C stands for chloramphenicol. </div> | ||
| − | + | <div id="apDiv22">2.The killing ability of the two warriors are not equal We calculate the ratio of warrior 1/warrior 2 in group 3 and group 6. If the ratios are alike, then the killing ability of the warriors are similar. Figure 2 shows the result.We can see that in the medium containing A+K, the amount of the warriors from two sides are roughly equal. At the same time, in the medium containing C, the amount of warrior 2 is much larger.This is easy to explain as not only does warrior 2 kills warrior 1, but warrior 1 kills itself as well. *A+K stands for ampicilin+kanamycin,C stands for chloramphenicol. </div> | |
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| − | <div id="apDiv22">2.The killing ability of the two warriors are not equal We calculate the ratio of warrior 1/warrior 2 in group 3 and group 6. If the ratios are alike, then the killing ability of the warriors are similar. Figure 2 shows the result.We can see that in the medium containing A+K, the amount of the warriors from two sides are roughly equal. At the same time, in the medium containing C, the amount of warrior 2 is much larger.This is easy to explain as not only does warrior 2 kills warrior 1, but warrior 1 kills itself as well. *A+K stands for ampicilin+kanamycin,C stands for chloramphenicol. </div> | + | |
<div id="apDiv21">IV.Results: </div> | <div id="apDiv21">IV.Results: </div> | ||
<div id="apDiv24">B.Test if the killing ability is adequate <br> | <div id="apDiv24">B.Test if the killing ability is adequate <br> | ||
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In contrast with the result above, we find that when we mix beggar1 and warrior 2 together, the amount of warrior2 surely surpasses that of beggar1. Besides, the killing ability of warrior2 is adequate. The killing ability is neither too strong when the ratio is 1:1, nor is it too weak when the ratio is 1:10. <br> | In contrast with the result above, we find that when we mix beggar1 and warrior 2 together, the amount of warrior2 surely surpasses that of beggar1. Besides, the killing ability of warrior2 is adequate. The killing ability is neither too strong when the ratio is 1:1, nor is it too weak when the ratio is 1:10. <br> | ||
Furthermore, we compared the amount of beggars when they are cultured alone with the amount of beggars when they are cultured with warriors. We calculated the amount of beggars in group 7/group 8 to see the change in beggar1 and in group11/group12 to see the change in beggar2.From figure 4, we can see that the amount of beggar1 drops drastically when it is cultured with warrior2, which means that warrior2 really work. As the amount of beggar2 does not have an apparent change, we may say that warrior1’s killing ability is surely too weak. </div> | Furthermore, we compared the amount of beggars when they are cultured alone with the amount of beggars when they are cultured with warriors. We calculated the amount of beggars in group 7/group 8 to see the change in beggar1 and in group11/group12 to see the change in beggar2.From figure 4, we can see that the amount of beggar1 drops drastically when it is cultured with warrior2, which means that warrior2 really work. As the amount of beggar2 does not have an apparent change, we may say that warrior1’s killing ability is surely too weak. </div> | ||
| − | <div id="apDiv26"><img src="fig1. | + | <div id="apDiv26"><img src="https://static.igem.org/mediawiki/2017/5/5a/Tsinghua-a-killing-fig1.jpg" width="407" height="299"></div> |
| − | <div id="apDiv27"><img src="fig2. | + | <div id="apDiv27"><img src="https://static.igem.org/mediawiki/2017/b/b9/Tsinghua-a-killing-fig2.jpg" width="380" height="303"></div> |
| − | <div id="apDiv28"><img src="fig3. | + | <div id="apDiv28"><img src="https://static.igem.org/mediawiki/2017/f/f1/Tsinghua-a-killing-fig3.jpeg" width="399" height="545"> <img src="https://static.igem.org/mediawiki/2017/5/52/Tsinghua-a-killing-fig4.jpeg" width="376" height="297"></div> |
<div id="apDiv30">V.Analysis:</div> | <div id="apDiv30">V.Analysis:</div> | ||
<div id="apDiv31"> Firstly, we ensure that our warriors can really kill. The amount of beggars do decrease when the warriors from the other side is added and the more warriors, the less beggars.(figure 3) Secondly,we get to the conclusion that the killing ability of the two warriors are apparently not equal. The killing ability of warrior2 is actually quite ideal as it does not kill itself and that its killing ability towards beggar1 is obvious but not excessively strong. The killing ability of warrior1 is rather weak. The weakness is partly because that warrior1 kills itself, and partly because it does not effectively kill beggar2. This result is consistent when our orthogonality test, as the orthogonality of warrior2 is quite well, but the orthogonality of warrior1 is not very good. In order to solve this problem, we have to modify our gene circuit so that warrior 1 does not kill itself and at the same time, effectively kill beggar2. </div> | <div id="apDiv31"> Firstly, we ensure that our warriors can really kill. The amount of beggars do decrease when the warriors from the other side is added and the more warriors, the less beggars.(figure 3) Secondly,we get to the conclusion that the killing ability of the two warriors are apparently not equal. The killing ability of warrior2 is actually quite ideal as it does not kill itself and that its killing ability towards beggar1 is obvious but not excessively strong. The killing ability of warrior1 is rather weak. The weakness is partly because that warrior1 kills itself, and partly because it does not effectively kill beggar2. This result is consistent when our orthogonality test, as the orthogonality of warrior2 is quite well, but the orthogonality of warrior1 is not very good. In order to solve this problem, we have to modify our gene circuit so that warrior 1 does not kill itself and at the same time, effectively kill beggar2. </div> | ||
Revision as of 16:33, 21 October 2017
TSINGHUA-A