Liu Zichen (Talk | contribs) |
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<h3>After construction</h3> | <h3>After construction</h3> | ||
− | <p>After the PCR product was harvested then cleavage:<br>Cutsmart® buffer 5μl<br>XbaI 1μl< | + | <p>After the PCR product was harvested then cleavage:<br>Cutsmart® buffer 5μl<br>XbaI 1μl<br>SacI 1μl<br>PCR product 43μl<br>After overnight digestion with the linearized plasmid pEX18 for 1 hr |
− | + | ||
Transformation of the large intestine<br>PxylA-xylR was cleaved by SacI on plasmid pAX01<br>The pEX18-lys, which has been added with lys, was linearized by SacI single digestion<br>The two are connected after 1hr<br>Transform into E.coli</p> | Transformation of the large intestine<br>PxylA-xylR was cleaved by SacI on plasmid pAX01<br>The pEX18-lys, which has been added with lys, was linearized by SacI single digestion<br>The two are connected after 1hr<br>Transform into E.coli</p> | ||
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<a href="#pic_fif"> | <a href="#pic_fif"> | ||
<img src="https://static.igem.org/mediawiki/2017/5/57/1234qaz%E5%89%AF%E6%9C%AC.png"></a> | <img src="https://static.igem.org/mediawiki/2017/5/57/1234qaz%E5%89%AF%E6%9C%AC.png"></a> | ||
− | <div align="center"><p style="font-size: | + | <div align="center"><p style="font-size:1.1rem;text-align:center"><br/>E.coli after transformation</p></div> |
</div> | </div> | ||
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<a href="#pic_sixteen"> | <a href="#pic_sixteen"> | ||
<img src="https://static.igem.org/mediawiki/2017/7/76/Qazwsx.png"></a> | <img src="https://static.igem.org/mediawiki/2017/7/76/Qazwsx.png"></a> | ||
− | <div align="center"><p style="font-size: | + | <div align="center"><p style="font-size:1.1rem;text-align:center"><br/>the data provided to iGEM-Tianjin for reference</p></div> |
</div> | </div> | ||
Revision as of 02:38, 26 October 2017
/* OVERRIDE IGEM SETTINGS */
Helping TUST prepare for their first year of competition.
To begin with, we gave them construct suggestions, communicated with their team adviser. In 24/3/2017, we were invited to TUST to carried out recruiting propaganda for them, and provided our construction guide.
2017.08.01
In 2017.08.01, TUST and Tianjin held a meeting together, during the meeting, TUST gave an account of their recent progress, we suggested them to add some new synthetic routes and elements, and probably new strain instead of simply Xylinus for fibrin.
Later on
Later on, TUST came again with their modeling problem, we gave them suggestion that we can share the Fluorescence method and modeling method with them, design Fluorescence experience and modeling together since it’s their first year competition.
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)
Upper Primer:gcTCTAGAATGAAATACCTGCTGCCGAC
Lower 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
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
Test the mini system for OUC
Mini system is a system composed of a mini promoter (about 100bp) and a mini terminator, and it is applicable for yeast. Because of the similarity of our biological chassis, they reached out to us and wanted us to run some control experiments for them. Using the same protocol and their mini system, we need to transform the system into our Saccharomyces cerevisiae in order to test if the plasmid will still function the same.
Protocol
2. Protocol: initial OD600 = 0.01 Test OD600 and threshold fluorescence every other 5-6 hours Exciting light λ=502/Emissive light λ=532
Result
This is our testing result for team OUC, turns out that our results reflects the same conclution. Thanks OUC for thiet trust and we are so glad that we helped.
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.
This one goes deeper
A good rule for rocket experimenters to follow is this: always assume that it will explode. The vehicle explodes, literally explodes, off the pad. The simulator shakes you a little bit, but the actual liftoff shakes your entire body and soul.
Even deeper
The path of the righteous man is beset on all sides by the iniquities of the selfish and the tyranny of evil men. Blessed is he who, in the name of charity and good will, shepherds the weak through the valley of darkness, for he is truly his brother's keeper and the finder of lost children.
And a level-3 title again
It has been said that astronomy is a humbling and character-building experience. There is perhaps no better demonstration of the folly of human conceits than this distant image of our tiny world. To me, it underscores our responsibility to deal more kindly with one another, and to preserve and cherish the pale blue dot, the only home we've ever known.