Tochingyuet (Talk | contribs) |
|||
Line 37: | Line 37: | ||
<p><h2>Mentoring with UCCKE</h2></p> | <p><h2>Mentoring with UCCKE</h2></p> | ||
<p style="font-family: roboto;font-size:115%;"> | <p style="font-family: roboto;font-size:115%;"> | ||
− | The Hong Kong UCCKE team aims to create biobricks that detect and control gout by degrading uric acid and transport it. The Hong Kong UCCKE team presented their idea to us and we proposed that they can run an assay with plate reader measuring the fluorescence signal of the detection module, and incubating transformed E. coli in LB medium with saturated uric acid over night for the assay of the degradation module. Detailed protocols can be viewed | + | The Hong Kong UCCKE team aims to create biobricks that detect and control gout by degrading uric acid and transport it. The Hong Kong UCCKE team presented their idea to us and we proposed that they can run an assay with plate reader measuring the fluorescence signal of the detection module, and incubating transformed E. coli in LB medium with saturated uric acid over night for the assay of the degradation module. Detailed protocols can be viewed below. |
We have visited each others lab and presented our project in July, and we have suggested them to amplify the synthesized DNA from IDT for stock and use screening method other than restriction digestion(e.g TA clone) as the insert and vector have similar size. We also pointed the need of negative and positive controls in their assays. | We have visited each others lab and presented our project in July, and we have suggested them to amplify the synthesized DNA from IDT for stock and use screening method other than restriction digestion(e.g TA clone) as the insert and vector have similar size. We also pointed the need of negative and positive controls in their assays. | ||
− | Apart from providing advice and analysing the results, we also provided instrument for the standard cloning of their biobricks. As one of the secondary schools with advanced lab settings, they still lacks important devices like -80C fridges for storing competent cell. Thus we helped them to clone biobricks in parallel. We also helped preparing the sequencing of their plasmids and perform the uric acid assay through plate reader for | + | Apart from providing advice and analysing the results, we also provided instrument for the standard cloning of their biobricks. As one of the secondary schools with advanced lab settings, they still lacks important devices like -80C fridges for storing competent cell. Thus we helped them to clone biobricks in parallel. We also helped preparing the sequencing of their plasmids and perform the uric acid assay through plate reader for their referece. |
In return, the Hong Kong UCCKE team realised our hardware detection kit by translating our blueprint into 3d model and printed it for us. They also provided us suggestions on the improvement of the prototype regarding 3d printing production. The resulting product is as shown in the following figures.</p> | In return, the Hong Kong UCCKE team realised our hardware detection kit by translating our blueprint into 3d model and printed it for us. They also provided us suggestions on the improvement of the prototype regarding 3d printing production. The resulting product is as shown in the following figures.</p> | ||
Line 47: | Line 47: | ||
<p style="font-family: roboto;font-size:115%;"> | <p style="font-family: roboto;font-size:115%;"> | ||
The Hong Kong HKUST team aspire to create a safety switch that would not compromise the functionality of original biobrick, but able to revert the transformant back to wild-type with time. We helped HKUST team’s progress by providing them a Cython-implemented Python interface. It enables them to use the ViennaRNA package functions(written in C language) in Python interface, so they can easily model the dissociation constant for the antisense mRNA and transcript mRNA. | The Hong Kong HKUST team aspire to create a safety switch that would not compromise the functionality of original biobrick, but able to revert the transformant back to wild-type with time. We helped HKUST team’s progress by providing them a Cython-implemented Python interface. It enables them to use the ViennaRNA package functions(written in C language) in Python interface, so they can easily model the dissociation constant for the antisense mRNA and transcript mRNA. | ||
− | We also help characterizing their biobricks by running an GFP assay, | + | We also help characterizing their biobricks by running an GFP assay, detailed protocol and results attached below. In return, they provided us two Python scripts for our modelling of free energies of opened and closed toehold switches, by plotting ODE-based model and stochastic model using Euler’s method and Gillespie algorithm respectively. This facilitates our screening of optimal toehold switches in our program.</p> |
− | <img src="https://static.igem.org/mediawiki/2017/5/5d/CUHK_UST.jpg" width="450px" height="auto"> | + | <center> <img src="https://static.igem.org/mediawiki/2017/5/5d/CUHK_UST.jpg" width="450px" height="auto"> </center> |
− | + | <br> | |
− | + | <center> <img src="https://static.igem.org/mediawiki/2017/3/39/CUHK_collab_result.jpg" width="900px" height="auto"> </center> | |
+ | <br> | ||
+ | <center> <p>The test result of the characteriation of 6 biobricks provided by Hong Kong HKUST team. The biobricks were incubated with varying concentrations of AHT and measured the GFP fluorescence with COSTAR plate reader. Only pSB1C3_BBa_T9002 shows a positive trend with increasing AHT concentration. </p> | ||
<br> | <br> | ||
Line 70: | Line 72: | ||
<br><br> | <br><br> | ||
<b>HKUST:</b><br> | <b>HKUST:</b><br> | ||
− | The protocol is provided by HKUST here.</p> | + | The protocol is provided by HKUST <a href="https://static.igem.org/mediawiki/2017/a/a1/Hong_Kong_HKUST_sensing_characterization.pdf">here</a>. </p> |
</section> | </section> |
Revision as of 11:15, 30 October 2017