Difference between revisions of "Team:Tianjin/Collaborations"
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<h2>Helping TUST prepare for their first year of competition.</h2> | <h2>Helping TUST prepare for their first year of competition.</h2> | ||
| − | <p>To begin with, we | + | <p>To begin with, we offered them some constructive suggestions, and communicated with their team adviser. In 24/3/2017, we were invited to TUST to carried out a recruiting propaganda for them, and helped them form a team.</p> |
<div id="middle" name="middle"> | <div id="middle" name="middle"> | ||
<h3>2017.08.01</h3> | <h3>2017.08.01</h3> | ||
| − | <p>In 2017.08.01, TUST and Tianjin held a meeting together, during | + | <p>In 2017.08.01, TUST and Tianjin held a meeting together, during which TUST gave an account of their recent progress, we suggested them to add some new synthetic routes and elements in their project, and probably a new strain instead of simply Xylinus for fibrin.</p> |
</div> | </div> | ||
| Line 281: | Line 281: | ||
<div id="two" name="two"> | <div id="two" name="two"> | ||
<h3>Later on</h3> | <h3>Later on</h3> | ||
| − | <p>Later on, TUST came again with their modeling problem, we | + | <p>Later on, TUST came to us again with their modeling problem, we gladly suggested them to share the Fluorescence method and modeling method with us. Since it's their first year competition, we offered them to design Fluorescence experience and modeling together. </p> |
</div> | </div> | ||
| Line 306: | Line 306: | ||
<h2>Helping NKU with their plasmid construction</h2> | <h2>Helping NKU with their plasmid construction</h2> | ||
| − | <p>What we did：<br>Helping NKU-iGEM construct their plasmid<br>The cleavage sites XbaI and SacI were added by PCR before and after lysase (lys)<br> | + | <p>What we did：<br>Helping NKU-iGEM construct their plasmid<br>The cleavage sites XbaI and SacI were added by PCR before and after lysase (lys)<br>Forward Primer：gcTCTAGAATGAAATACCTGCTGCCGAC<br>Reverse Primer：cGAGCTCtcaatgcgtttccataatagcagc</p> |
<div id="three" name="three"> | <div id="three" name="three"> | ||
Revision as of 08:50, 26 October 2017
/* OVERRIDE IGEM SETTINGS */
Helping TUST prepare for their first year of competition.
To begin with, we offered them some constructive suggestions, and communicated with their team adviser. In 24/3/2017, we were invited to TUST to carried out a recruiting propaganda for them, and helped them form a team.
2017.08.01
In 2017.08.01, TUST and Tianjin held a meeting together, during which TUST gave an account of their recent progress, we suggested them to add some new synthetic routes and elements in their project, and probably a new strain instead of simply Xylinus for fibrin.
Later on
Later on, TUST came to us again with their modeling problem, we gladly suggested them to share the Fluorescence method and modeling method with us. Since it's their first year competition, we offered them to design Fluorescence experience and modeling together.
Helping NKU with their plasmid construction
What we did：
Helping NKU-iGEM construct their plasmid
The cleavage sites XbaI and SacI were added by PCR before and after lysase (lys)
Forward Primer：gcTCTAGAATGAAATACCTGCTGCCGAC
Reverse Primer：cGAGCTCtcaatgcgtttccataatagcagc
After construction
After the PCR product was harvested then cleavage：
Cutsmart® buffer 5μl
XbaI 1μl
SacI 1μl
PCR product 43μl
After overnight digestion with the linearized plasmid pEX18 for 1 hr
Transformation of the large intestine
PxylA-xylR was cleaved by SacI on plasmid pAX01
The pEX18-lys, which has been added with lys, was linearized by SacI single digestion
The two are connected after 1hr
Transform into E.coli
E.coli after transformation
Helped us to determine the fluorescence intensity of pRS416-CUP1p-RFP without copper induction in Saccharomyces cerevisiae BY4742 and BY4741.
Cultures were incubated overnight in SC-URA medium
Take the bacteria to the new SC-URA to adjust the OD600 value to 0.1 for 24 hours
Fluorescence was measured
Excitation 472nm
Radiation 532nm
In the case of
the data provided to iGEM-Tianjin for reference
Test the mini system for OUC
What we did：
To verify whether the system built by China Ocean University is still applicable in other species of Saccharomyces cerevisiae. For this reason, we used our laboratory-specific Saccharomyces cerevisiae with synthetic chromosome 10 to observe its fluorescence values.
Protocol：
Yeast with plasmid was incubated overnight in YPD + G418 medium
Transfer the bacteria to the new YPD + G418 and adjust the OD to 0.1
After incubation for 20 hours, the fluorescence was measured
Excitation light 502nm
Emitting light 532nm
The OD600 values were measured after fluorescence measurements
After comparison with the data provided by Ocean University, it is found that the experimental results are consistent, but the slight deviation of the data shows that the mini system has similar expression in different laboratories and yeast strains.
Testing results for OUC
What we ask OUC to do
Easy - to - error PCR library development. For our use of the CUP1 promoter error-prone PCR, amplification of our existing error-prone PCR library.
Specific steps:
1.Error-prone PCR
2. digestion
3. Purification / Adsorption
4. Connect
5. Large intestine transformation
Easy-to-error PCR protocol (100μl)：
5X buffer（140mM MgCl2, 250mM KCl, 50mM Tris, and 0.1%(wt/vol) gelatin）20μl
Template (iGEM-Tianjin provided) 4μl
Primers (iGEM-Tianjin provided) 4μl*2
10X dNTP (2mM dGTP, 2mM dATP, 10mM dCTP, and 10 mM TTP) 10μl
Taq polymerase 2μl
5mM MnCl2 10μl
ddH2O 46μl
94℃ 3min
94℃ 30s
53℃ 30s
72℃ 30s
72℃ 1min
recycle for 35 times
The PCR products were recovered and digested with BamHI and XbaI, and ligated with vector pRS416.
In the case of
After intestine turn to collect bacteria and plasmids
Sent a Collaboration Request and construct an alliance to build a worldwide database.
We came up with the idea that we could gather all the iGEM team whose project is about water pollution treatment together and build an alliance to unite all our social impact, knowledge and geographical advantages.
2017.08.12
2. In 12th of August, we had a voice conferencing with SJTU/SCUT/XMU/UCAS/JLU/FAFU, we discussed about how we want to use this alliance and came up with 2 conclusion: 1. Build a worldwide database containing contents of heavy metals in soil 2. Mutual improve our social impact
2017.08.23
In 23th of August, we sent a Collaboration Request to iGEM official website, the next day Ana Sifuentes replied our message and posted our request in the iGEM official website,
and we received the response of many teams like team EXETER and team CSMU NCHU TAIWAN, with their information, we construct a database which contains the worldwide data of contents of Cu2+/Cd2+ in soil or water. And we build it based on map of the world. We received team CSMU NCHU TAIWAN’s kindly help, they offered us information about major metal pollution incidents in Taiwan as well as the real time monitoring data of places that have the potential to outbreak serious pollution incidents oneday.
Helping CQU construct their team and sign up for their first year
We offered team CQU China many constructing advice including finding team advisor and allocating funds. Next yeat will be their first year joining iGEM, and we will continue helping them until they are ready for their Gamboree. Hope their result in next year will be great!
Filmed a biosafety video together with other 12 teams
12 teams gather together to film a biosafety video, every team took different topics, but all based on Yale biosafety manual.
