iGEM 2014 — Team UESTC-China

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# Team:UESTC-China/Notebook

### From 2014.igem.org

UESTC-China

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[Home](https://2014.igem.org/Team%3AUESTC-China)

Team

[* Members](https://2014.igem.org/Team%3AUESTC-China/Team)
[* Attribution](https://2014.igem.org/Team%3AUESTC-China/Attribution)
[* Team Profile](https://igem.org/Team.cgi?year=2014&team_name=UESTC-China)
[* Notebook](https://2014.igem.org/Team%3AUESTC-China/Notebook)

Project

[* Overview](https://2014.igem.org/Team%3AUESTC-China/Project)
[* Design](https://2014.igem.org/Team%3AUESTC-China/Design)
[* Result](https://2014.igem.org/Team%3AUESTC-China/result)
[* Future work](https://2014.igem.org/Team%3AUESTC-China/Futurework)
[* Judging Criteria](https://2014.igem.org/Team%3AUESTC-China/Judingform)

Modeling

[* Overview](https://2014.igem.org/Team%3AUESTC-China/Modeling)
[* HPS/PHI](https://2014.igem.org/Team%3AUESTC-China/Modeling1)
[* FALDH/FDH](https://2014.igem.org/Team%3AUESTC-China/Modeling2)
[* AtAHA2](https://2014.igem.org/Team%3AUESTC-China/Modeling3)

Parts

[* BioBrick](https://2014.igem.org/Team%3AUESTC-China/BioBrick)
[* Protocol](https://2014.igem.org/Team%3AUESTC-China/Protocol)

Human Practice

[* Questionnaire](https://2014.igem.org/Team%3AUESTC-China/Questionnaire.html)
[* Knowledge quiz](https://2014.igem.org/Team%3AUESTC-China/Knowledgequiz)
[* Lecture](https://2014.igem.org/Team%3AUESTC-China/Lecture.html)
[* Communication](https://2014.igem.org/Team%3AUESTC-China/Communication.html)
[* Art](https://2014.igem.org/Team%3AUESTC-China/Art)

Safety

[* Safety](https://2014.igem.org/Team%3AUESTC-China/Safety)

# Notebook

* February

  **Feb 14**
  iGEM 2014 registration opens.
  **Feb 20**
  Site: West 304 of main building in UESTC
  Team of iGEM 2014 in UESTC is founded.
  **Feb 21**
  Site: West 304 of main building in UESTC
  We name our team Green Life;
  Discuss the direction we will focus on for the iGEM 2014.
  **Feb 24**
  Site: West 304 of main building in UESTC
  We meet the supervisors Yong Zhang, Xuelian Zheng, Lixia Tang, discussing the direction and acquiring their opinions of our project.
* March

  **March 3**
  Site: West 304 of main building in UESTC
  We firstly choose plant as our receptor, each member of our team give advices for the details we plan to do, such as which plant to use, tobacco, arabidopsis or any other plant?
  **March 10**
  Site: West 304 of main building in UESTC
  We talk with our supervisors and decide to use tobacco as our receptor for its simpleness as model organism.
  **March 17**
  Site: West 304 of main building in UESTC
  We discuss the project and firstly decide to create a plant which can absorb formaldehyde largely by synthetic biology. And we choose Yasong Cui as our team leader.
  **March 24**
  Site: West 304 of main building in UESTC
  Team leader assign task for us;
  The first phase for our task begins, searching references and designing our logo.
  **March 31**
  We register our team with the name of Green Life;
  iGEM 2014 registration closes;
  Team registration fee due.
* April

  **April 9**
  Site: West 304 of main building in UESTC
  Group meeting for the references and information we have researched last few days. Team leader Yasong Cui and Jingyao Li report the key enzymes through the formaldehyde metabolic pathways. *HPS*, *PHI* are critical enzymes of RuMP pathway in formaldehyde assimilation. The references show that the transformation method is chloroplast transformation. The crucial enzyme in amplifying stomas of foliage is *AHA2*.
  **April 13**
  Site: West 304 of main building in UESTC
  Group meeting for the reference reading.
  Mingyuan Wang reports one gene (*AdCP*) of pollen sterility;
  Jiao He reports *FDH* which can amplify metabolic pathway in plant;
  Yasong Cui reports gene *AHA2* which can amplify stoma of foliage.
  **April 16**
  Site: West 304 of main building in UESTC
  Group meeting for the wiki constructing;
  The responsible person of wiki, Zhiqiang Yan assigns tasks for the rest.
  **April 20**
  Site: West 304 of main building in UESTC
  Jie Li reports *FALDH* which can amplify metabolic pathway in plant;
  Rui Liu reports some information about *HPS*, *PHI*;
  Yasong Cui reports some information about *AHA2*.
  **April 23**
  Site: West 304 of main building in UESTC
  Vector construction and confirmation of the gene (*HPS*, *PHI*, *FDH*, *FALDH*, *AHA2*, *AdCP*) being used in our project.
  **April 23**
  Site: online
  We change the team name Green Life into UESTC-China.
  **April 25**
  Start to design a questionnaire about iGEM and our project. And plan to do a small range research.
  **April 30**
  Site: The seventh middle school in Chengdu
  We introduce the concept and racing type of iGEM to the students in the seventh middle school who were the best students in Sichuan province.
  Site: West 304 of main building in UESTC
  We talk about the PrbcS-3C promoter sequence and the gene sequence of transit peptide of chloroplast.
* May

  DNA Distribution Kit sent to teams
  **May 01**
  iGEM 2014 Late registration closes
  **May 3**
  Site: West 304 of main building in UESTC
  Decision on the backbone vector.
  The backbone named pXZY006 is proffered by one of our advisor Zhengyang Xie. And we designate it piGEM001.
  Weixin Liu finds some chloroplast and cytoplasm transit peptide from Uniprot database.
  **May 7**
  Site: West 304 of main building in UESTC
  Take a group meeting and design the vectors we plan to use.
  **May 14**
  Site: West 304 of main building in UESTC
  Group meeting.
  We discuss some details in the designing of vector constructing;
  At the end of the meeting, we started our experiments.
  **May 19**
  Site: West 121 of main building in UESTC
  We extract pXZY006 plasmid from bacterium culture using TIANGEN kit.
  Using SpeⅠand HpaⅠto do the double digestion of pXZY006 and F01 (contains *HPS* ) and do the ligation using T4 DNA ligase.
  **May 22**
  Site: West 121 of main building in UESTC
  Do the transformation in competent recipient *E.coli* cell.
  **May 23**
  Site: West 121 of main building in UESTC
  Repeat the experiments we do in May 19 and May 22 for the outcome is not that satisfactory.
  Also we do not use the T4 ligase to do the ligation but by Gibson assembly.
  **May 28**
  Site: West 121 of main building in UESTC
  Do the colony PCR to testify if the F01 has ligated to pXZY006.
  And it comes out with positive clones, so the vector piGEM001 has been successfully constructed.
  **May 29**
  Site: West 304 of main building in UESTC
  Group meeting;
  Decide to build the second vector piGEM002.
  Ligate F05, F02, and F03 to piGEM001 to construct piGEM002;
  Ligate F06, F07 to piGEM002 to construct piGEM011.
  **May 30**
  Site: West 121 of main building in UESTC
  Ligate F06 and F07 to iGEM001.
  **May 31**
  Site: West 121 of main building in UESTC
  Do the transformation using heat shock.
  The outcome is satisfactory that there is no clone in control group and thirteen clones in experimental group.
* June

  **June 5**
  Site: West 121 of main building in UESTC
  Do the colony PCR to testify if the F06 and F07 have ligated to piGEM001.
  This ligation fail due to the outcome of colony PCR.
  **June 6**
  Site: West 121 of main building in UESTC
  Repeat the experiments we did on May 31.
  **June 7**
  Site: West 121 of main building in UESTC
  Do the transformation and colony PCR.
  The outcome of colony PCR still demonstrate we fail again.
  **June 8-13**
  Site: West 121 of main building in UESTC
  We do the ligation and transformation repeatedly, asking help from our advisors, finally we got the right outcome of piGEM011 (not the originally piGEM011 we planned to construct, used to be F06+F07+piGEM002 and now is F06+F07+piGEM001).
  **June 14**
  Site: West 121 of main building in UESTC
  We ligate F02, F03, and F05 but fail.
  **June 15**
  Site: West 121 of main building in UESTC
  We ligate F02, F03, and F05 but fail.
  **June 16**
  Site: The area before Sun Shine dining hall in UESTC
  We make a knowledge quiz which contains a few questions concerning iGEM or synthetic biology to expand people's horizon.
  **June 19**
  Open Office Hours -- ask safety questions, meet other iGEMers!
  **June 23**
  About our Lab form due
  **June 17-30**
  Site: West 121 of main building in UESTC
  We do the multiple ligation of F02, F03, and F05 repeatedly for changing the parameters in experiments but fail.
* July

  **July 1-16**
  Site: West 121 of main building in UESTC
  We do the multiple ligation of F02, F03, and F05 repeatedly for changing the parameters in experiments but fail. And finally we abandon F03 and F05, ‘cause no matter how we improve the criteria of our experiments we still get the fail result. But we still add F02 into piGEM001 to construct the backbone.
  **July 13**
  Site: West 121 of main building in UESTC
  Start to construct DNA samples in pSB1C3.
  **July 16**
  Site: West 121 of main building in UESTC
  We ligate F02 to piGEM001 as the new vector piGEM002.
  **July 21**
  Preliminary safety forms due.
  **July 23**
  Site: West 121 of main building in UESTC
  Group meeting.
  We decide to construct the rest vectors piGEM003, piGEM004, piGEM005, piGEM006, piGEM007, piGEM008, piGEM009, piGEM010.
  **July 25**
  Track selection due;
  Site: West 121 and west 111 of main building in UESTC
  We communicate with the students from SCU. We exchange our project to each other and make a long talk about what we have done during the last month.
  **July 26**
  Site: West 121 and west 111 of main building in UESTC
  From July 25th, we start to do the transformation part. That really requires large volume of work. Four members of our team do the most of the work,and other members also help a lot.
  **July 27**
  Site: Before the main building of UESTC
  The members of our team gather and take a photo.
  **July 28**
  Site: West 121 of main building in UESTC
  Start to construct the vector piGEM003, piGEM004, piGEM005, piGEM006, piGEM008, piGEM010.
  **July 29**
  Site: West 121 of main building in UESTC
  Go on revising the questionnaire about iGEM. And plan to start a second research.
  **July 28-31**
  Site: West 121 of main building in UESTC
  Go on constructing the vector piGEM003, piGEM004, piGEM005, piGEM006, piGEM008, piGEM010.
  Start to construct piGEM007.
* August

  **August 2**
  Site: West 121 of main building in UESTC
  Start to construct the vector piGEM009.
  Do the colony PCR in the eight vectors to detect if the colony is positive or not.
  **August 3**
  Site: West 121 of main building in UESTC
  Amplify 6\*A-MASS, GSG-P2A, GSG-T2A, GSG-F2A by PCR.
  **August 5**
  Site: West 121 of main building in UESTC
  Amplify *AtAHA2* by PCR.
  **August 8**
  Site: The playground of UESTC and the opening area in front of supermarket
  We go to the playground and do the research for our questionnaire.
  **August 11**
  Site: West 121 of main building in UESTC
  All the vectors (piGEM003-010) we planned to build have finished.
  **August 15**
  Team project descriptions due.
  **August 18**
  Site: West 206 of main building in UESTC
  During the summer vacation, besides communicating with the students in SCU, there was another activity which worth a notice, our principal came to our laboratory and gave us a speech, which totally stimulated us to work harder and excitedly. The principal told us do not be scared to have the new idea, 'cause the study we received today is to make us to speak out a new idea no matter it is wizard or not.
  **August 20**
  Site: West 121 of main building in UESTC
  For about two months work, our wiki has its fundamental appearance.
  **August 27**
  Site: West 304of main building in UESTC
  Group meeting for the progress of our project.
  **August 30**
  Site: Qingshui campus of UESTC
  To improve the outcome of our iGEM, we start to do a third research for our questionnaire today. 'cause the people from various places gather in our school for accompanying their children to go to school.
* September

  **September 1 to now**
  Site: West 121 of main building in UESTC
  On September 1st, when the eighth leaves of the first batch of tobacco seedlings appear, we start the formal experiments of formaldehyde detection. Before that, some pre-tests are made based on some corresponding references. Finding the right amount of formaldehyde is the hardest part. By means of a series of experiments, we determine the final protocol which includes the qualitative detection and quantitative detection. From the experimental results, we can draw the conclusion that our super plants have remarkable formaldehyde tolerance and can dramatically reduce the concentration of formaldehyde in the air.
  **September 1**
  Final safety forms due.
  **September 5**
  Giant Jamboree attendance fee due at 11:59PM EDT;
  Team rosters due.
  **September 6**
  Site: West 213 of main building in UESTC
  Extract the DNA and RNA from transgenis plant, testifying.
  Team rosters due.
  **September 7-15**
  Site: West 121 of main building in UESTC
  Sum up the information of the questionnaire, and draw up the statistics.
  **September 20-30**
  Site: West 121 of main building in UESTC
  Analyze the statistics we get from the questionnaire. And start to consider how to present the conclusions on the wiki.
  **September 24**
  Site: West 121 of main building in UESTC
  The DNA samples in pSB1C3 have been completely finished.
  **September 25**
  Site: West 121 of main building in UESTC
  In the beginning of this new semester, our school held a science and culture activity. We caught this opportunity to advertise iGEM and our ultimate products, the plants which can absorb formaldehyde. The leaders and many teachers in our school also participated this activity. We displayed the plants which already owned the several genes which worked in the absorption of formaldehyde, to the people who visited our project. And the people visited our plant all said they want to owe one if there is already market available.
* October

  **October 1**
  Site: West 121 and west 111 of main building in UESTC
  Examine the formaldehyde tolerance of the transgenis plant and wildtype plant.
  **October 5**
  Site: West 121 and west 111 of main building in UESTC
  Examine the formaldehyde tolerance of the transgenis plant and wildtype plant.
  **October 3-8**
  Site: West 121 and west 111 of main building in UESTC
  Improve the outcomes of Questionnaire researching.
  **October 3-8**
  Site: West 121 and west 111 of main building in UESTC
  Keep on constructing our wiki and doing the examinations of our super plant's tolerance of formaldehyde.
  **October 9 to now**
  Site: West 121 and west 111 of main building in UESTC
  Keep on constructing our wiki, doing the examinations of our super plant's tolerance of formaldehyde and extract DNA and RNA from transgenis tobacco.
  **October 10**
  Site: West 121 and west 111 of main building in UESTC
  We mimic the iGEM jamboree spot and to present our project to the teachers.
  **Up to October 17**
  We keep on constructing our wiki, examining the writing error and lack of standardization and replenishing the picture and results of our project.
  **Up to now**
  We keep on replenish our experiments, and examining the formaldehyde tolerance of transgenis plant. And continue to picture the differences of formaldehyde tolerance in transgenis and wildtype plant.

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