Team:ZJU-China/Notebook

Notebook

  • September
  • 9.5.2015

    Finished the overview of entire project.

  • 9.4.2015

    • Bacteria solution PCR: OrfX. One positive clone of 20 samples.

    • DNA sequencing: OrfX.

  • 9.2.2015

    Dynamic Light Scattering(DLS) experiment to measure the particle size of embedded E.coli.

  • 9.1.2015

    The sequence of OrfX was not correct because of the wrong template, thus we had to do the related experiments again.

  • August
  • 8.31.2015

    Took photos and prepared for the banner~



  • 8.30.2015

    The sequence of metK and tcdA1 were confirmed, which marked the success in standardization of 2 parts!!

  • 8.29.2015

    Bacteria solution PCR: metK and OrfX. Both had positive clones. DNA sequencing: metK and OrfX. T4 ligation and transformation: plu0840+pSB1C3, plu1537+pSB1C3, tcdA1+pSB1C3. Selected from single colonies: CDS-tcdA1.

  • 8.28.2015

    • Selected from single colonies of metK and OrfX.

    • Submit the safety form. Selected from single colonies: CDS-plu0840. Double enzyme digestion and DNA gel extraction: plu1537, tcdA1, plu0840. Plasmid extration and double enzyme digestion: RFP. Protein extraction and SDS-PAGE: tcdA1, plu1537, plu0840. T4 ligation and transformation: plu0840+pSB1C3, plu1537+pSB1C3, tcdA1+pSB1C3. Bacteria solution PCR: CDS-plu0840.

  • 8.27.2015

    Seamless assembly: plasmid+OrfX, plasmid+metK. PCR: CDS-tcdA1. Double enzyme digestion: RFP+plasmid. Seamless assembly: CDS-plu0840. PCR and DNA gel extraction: CDS-tcdA1. Changed pSB1A3 to pSB1C3: plu0840, plu1537, tcdA1. Transformed them into E.coli DH5α, selected from single colonies and used single enzyme digestion to verify the got plasmids.

  • 8.26.2015

    Seamless assembly: plasmid+CDS-tcdA1, plasmid+CDS-plu0840. DNA gel extraction: tcdA1. PCR: device-plu0840.

  • 8.25.2015

    The sequence of ermE+plasmid, Frr+plasmid and plu1537+plasmid were confirmed, which marked the success in standardization of 3 parts!!

  • 8.24.2015

    • Bacteria solution PCR: metK and OrfX. Only negative clone. Extraction of plasmid mCherry and double enzyme digestion. DNA sequencing: ermE+plasmid, Frr+plasmid, plu1537+plasmid. Selected from single colonies of plu1537.

    • Power cut. We played table tennis near our lab.


  • 8.23.2015

    • Selected from single colonies of metK and OrfX.

    • Transformed standard plu1537 plasmid into BL-21. Bacterium solution PCR: ermE and Frr.

    • We kept a Madagascar cockroach as our pet~




  • 8.22.2015

    • Selected from single colonies of ermE and Frr. Extraction of plasmid (standard plu1537).

    • Bacterium solution PCR: standard plu1537.

  • 8.21.2015

    • DNA sequencing. Seamless assembly: plasmid+1537, plasmid+tcdA1-L+ tcdA1-M+ tcdA1-R, plasmid+ermE, plasmid+Frr.

    • Successfully booked the HI-Boston Hostel and paid the full payment~ Seamless assembly: plasmid+metK, plasmid+OrfX.

  • 38th Group meeting

    8.20.2015

    38th Group meeting. Introduced a software named Mou which could enhance the efficiency of writing wiki. Discussed the business planning of our product and the photos on the wiki.

  • 8.20.2015

    • Extraction of plasmid mCherry and double enzyme digestion. Seamless assembly: plasmid+1537, plasmid+tcdB1. PCR: ermE, Frr, metK and OrfX. Clean up and DNA gel extraction.

    • Capsulated Streptomyces Avermitilis toxicity test. TT01 toxicity verification.

    • PCR: ermE, Frr, metK and OrfX. Clean up and DNA gel extraction.

    • Again used flow cytometry to check the diameter of the embedded E.coli and tried to count the E.coli which were successfully embedded. The results were not that ideal again.

  • 8.19.2015

    • PCR: ermE, Frr, metK and OrfX. Clean up and DNA gel extraction.

    • Finished the background of avermectin and toxic protein. Sent bacterium sample to BNU and we would continue to help them.


  • 8.18.2015

    • PCR: CDS tcdB1, CDS 1537, tcdA1-L, mcherry. DNA sequencing.

    • Half-done the model of degration of toxic protein. Got the kit for detection avermectin.

  • 8.17.2015

    • PCR: CDS tcdB1, CDS plu1537 and mCherry. DNA gel extraction: tcdA1-L and plu0840L. Seamless assembly: plasmid+pBad+plu1537.

    • Detected the relationship between distance of food source and food consumption.

    • Designed the primers for Frr, metK, OrfX and ermE again because the need for standardization.

  • 37th Group meeting

    8.16.2015

    37th Group meeting. Built the framework for wiki. Discussed the Syn-bio talks on next Saturday.

  • 8.16.2015

    • Preparation of competent cells.DNA gel extraction.

    • Verification of the bait attractiveness.

  • 8.15.2015

    • PCR: pBad, tcdA1-L, tcdB1, plu0840L and plu1537. Purchased E.coli BL21.

    • The experiment concerning Streptomyces avermitilis finished 80%.

    • Lovely poppy~


  • 8.14.2015

    • PCR and clean up. Concluded the successful PCR parameters of different gene clusters.

    • Test the Avermectin detection kit with pure Ivermectin.

    • Designed the primers for plu0840, plu1537, tcdA1 and tcdB1 again bucause of the need for standardization.


  • 8.13.2015

    • Overlap: pBad and tcdA1-L, pBad and tcd-B1, pBad and plu1537. Extraction of plasmid mCherry and enzyme digestion.

    • Completed wiki page for 2nd model. Drew the 3D ball-and-stick models for CNC with Solidworks. Completed the device for killing termites.

    • Yummy hotpot~

    • Completed the SCM program for HAZU and they affirmed our help.




  • 8.12.2015

    • Overlap: 0840L and 0840R.

    • Tried to purify the embedded E.coli with the E.coli which were able to express RFP.

    • Used flow cytometry to check the diameter of the embedded E.coli and tried to count the E.coli which were successfully embedded. The results were not that ideal.


  • 8.11.2015

    • PCR: pBad, tcdA1-L, tcdA1-M, tcdA1-R, tcdB1, 0840L, 0840R, 1537, GFP.

    • PCR: ligated product. Optimized the PCR program. Cleaned up. PCR again.

    • SEM scanned our samples to verify whether Streptomyces avermitilis were successfully embedded with CNC.

    • Fed the termites with Photorhabdus luminescens TT01 bacterium cells.


  • 8.10.2015

    • PCR: pBad, tcdA1-L and GFP.

    • Used Bgl2 and EcoR1 digested inserted fragment and cleaned up the product. Ligating fragment into plasmid with T4 ligase.

    • Specified the standardization requirements and laid the foundation for primer designing.

    • Mixed blue-stained termites with red-stained termites to verify trophallaxis.


  • 35th Group meeting

    8.9.2015

    35th Group meeting. Planned to hold a seminar for freshmen. Tried to design a device for attracting and killing termites.

  • 8.9.2015

    • PCR: tcdA1-M, tcdA1-R, tcdB1, plu0840L, plu0840R and plu1537.

    • Completed our framework and philosophy of safety.

    • Extracted the plasmid containing RFP from DH5α. PCR: RFP and confirmed by gel electrophoresis.


  • 8.8.2015

    Visited Shanghai Science&Technology Museum and promoted our Synbio-cards(Polypoly card).

  • 8.8.2015

    Visited Shanghai Science&Technology Museum and promoted our Synbio-cards(Polypoly card).

  • 8.7.2015

    One step cloning protocol finished. Designed the questionaire about termites.

  • 8.7.2015-8.8.2015

    PCR: pBad, GFP, tcdA1-M, tcdA1-R.

  • 34th Group meeting

    8.6.2015

    34th Group meeting. Communicated with 2 members of SJTU in our lab. Began to write wiki. Attached more importance to safety.

  • 8.6.2015

    • Drawing plasmid circuits in snapgene.(toxic protein)

    • Stained 50 termites in blue and 50 in red for trophallaxis experiments.


  • 8.5.2015

    • Discussed the circuit construction and experiments planning of toxic protein.

    • Decided to give up confocol because we couldn't find the specific dye for CNC.


  • 8.4.2015

    Completed the One Step Cloning protocol.

  • 33rd Group meeting

    8.2.2015

    33rd Group meeting. Began to build framework for wiki. Introduced the habits and characteristics of termites.

  • July
  • 32nd Group meeting

    7.30.2015

    32nd Group meeting. Planned to promote synthetic biology and our project in Shanghai Science & Technology Museum. Polished up the Synbio-cards(Polypoly card).

  • 7.28.2015-8.4.2015

    PCR: pBad, tcdA1-L, tcdA1-M, tcdA1-R, tcdB1, plu0840L, plu0840R, plu1537 and GFP.

  • 7.28.2015

    Giant Jamboree registration completed.

  • 7.26.2015

    • Listed 15 main problems we were confronted with, including safety, communication between dry group and wet groups.

    • Designed a new method to embed the bacterium with CNC.

  • 7.25.2015

    Designed the primers for plu0840, plu1537, tcdA1 and tcdB1 again bucause of the bug of extra RBS.

  • 7.24.2015

    Discussed helping BNU and HUST. Designed and alpha tested the Synbio-cards. Improved the modeling.

  • 7.19.2015-7.24.2015

    Attended NCTU meetup in Taiwan. Present our half-done project and communicated with other teams. Decided to help BNU and HUST.

  • 7.19.2015-7.24.2015

    Attended NCTU meetup in Taiwan. Present our half-done project and communicated with other teams. Decided to help BNU and HUST.

  • 7.18.2015

    • Listed and detailed the wiki requirements.

    • Fed the termites with Streptomyces Avermitilis bacterium cells.

  • 7.16.2015

    Prepared the home page and member page of our wiki. Prepared the poster for NCTU meetup.

  • 7.15.2015

    • Designed the primers for Frr, metK, OrfX and ermE again. Got our name card~

    • Used confocal to verify the successful package of E.coli with CNC. However, the result appeared not ideal and we couldn't find appropriate dye of CNC.


  • 7.14.2015

    Prepared for the souvenirs for NCTU meetup.

  • 31st Group meeting

    7.12.2015

    31st Group meeting. Reviewed the medal requirement and all the rules in lab.

  • 7.11.2015

    Mark-recapture method failed because we failed to recapture enough stained termites.

  • 7.3.2015-7.11.2015

    Exams of summer semester.

  • June
  • 30th Group meeting

    6.25.2015

    30th Group meeting. Planned to verify the package of CNC with confocal. Continued to prepare for the presentation and application scenario.

  • 6.24.2015

    • Completed 2 safety questionaires.

    • Prepared to learn confocol to verify the CNC package for E.coli.

  • 6.22.2015

    Put 150 blue stained termites back into the nest for mark-recapture method.

  • 29th Group meeting

    6.21.2015

    29th Group meeting. Discussed the application scenario for the 5000€ additional sponsership of SYNENERGENE.

  • 6.21.2015

    Contacted with Björn and he would help us animate the process of crystallization.
  • 6.20.2015

    TEM scanned our samples.

  • 6.19.2015

    Fed the termites with Photorhabdus luminescens TT01 bacterium solution to test the natural toxicity. Stained 150 termites in blue for future experiment.

  • 28th Group meeting

    6.18.2015

    28th Group meeting. 2nd modeling half done. Discussed the results of the SEM.

  • 6.17.2015

    TEM and SEM scanned our samples.

  • 6.17.2015

    TEM and SEM scanned our samples.

  • 6.15.2015

    Fed the termites with Streptomyces Avermitilis bacterium solution to test the natural toxicity.

  • 27th Group meeting

    6.14.2015

    27th Group meeting. Rearranged all the events with four quadrant method. Checked everything we have had.

  • 6.14.2015

    Solved the diffusion equation.


  • 6.13.2015

    Extracted the genome of Streptomyces avermitilis.

  • 6.12.2015

    Went to shanghai for the interview of visa. 12 members successfully passed the interview.

  • 26th Group meeting

    6.11.2015

    26th Group meeting. Discussed the half-done presentation and Skype conference with SYNENERGENE. Prepared for the visa.

  • 6.11.2015

    PCR: plu1537, with the template of the genome of TT01.
  • 6.10.2015

    • Paid for the air tickets.

    • Got the primers of toxic protein.

  • 6.8.2015

    Optimization of the model of termites finding food.





  • 25th Group meeting

    6.7.2015

    25th Group meeting. Polished up the previous modeling. Protocols about the experiments concerning bacterium. A member quit.

  • 6.7.2015

    Extracted the genome of TT01.
  • 6.6.2015

    Frozen the spores of Streptomyces avermitilis.

    Judge Haoqian Zhang from Peking University came to our lab. Frozen the spores of Streptomyces avermitilis.

  • 24th Group meeting

    6.4.2015

    24th Group meeting. Brainstormed the Synbio-cards and Synbio-workshops. Listed the datailed personal scheme about the time arrangement before summer holiday.

  • 6.3.2015

    • Discussed the characterization of E.coli embedded in CNC, including TEM, SEM, confocal, FTIR and flow cytometry.

    • Our proposal had been accepted and our team would be awarded financial support from SYNENERGENE! Cheers~

    • Brainstormed how to do human practice and use the €5000. We came up with the Synbio-cards and Synbio-workshops.

  • 6.2.2015

    Began to book the air tickets. Booked the first primers of toxic protein for one step cloning. Found the sequence of the gene we need in the Streptomyces avermitilis.

  • 6.1.2015

    SEM scanned our samples again. Debugged the first modeling.

  • May
  • 23rd Group meeting

    5.31.2015

    23rd Group meeting. Discussed the half-done modeling.

  • 5.31.2015

    Built up the model of termites finding food.
  • 5.31.2015

    Taught high school students to do experiment to extract DNA from epithelial cell of mouth.




  • 5.29.2015

    • Repeated the cellulase activity detection experiment. Meanwhile we measured the lysozyme activity in different parts of termites guts.

    • 22nd Group meeting. Specified the medal requirement. Began to prepare for the NCTU meetup.

    • SEM scanned our samples.

  • 5.28.2015

    Completed a model of a cartoon termite holding a sword. Cleaned up our lab and rearranged all the experiment materials we would need.

  • 5.26.2015

    Got the solid CNC with lyophilization. Built the framework of the first web page.

  • 5.25.2015

    Learned the skills of culturing Streptomyces avermitilis in the professor's lab.

  • 21st Group meeting

    5.24.2015

    21st Group meeting. Began to prepare for the presentation.

  • 5.22.2015

    iGEM Shanghai Tour. Took part in the conference held in NYU. Visited FDU and communicated with them.

  • 5.21.2015

    Chosen the certain toxic protein which we would transfer into E.coli: tcdA1, tcdB1,plu0840 and plu1537.

  • 5.21.2015

    20th Group meeting. Discussed the medal requirements and collaboration among groups.
  • 5.20.2015

    • Finished the protocol about TT01.

    • Measured the cellulase activity in termites head, forgut, midgut and hindgut.

  • 5.19.2015

    Detailed the medal requirements depending on our project. Finished the protocol about Streptomyces avermitilis.



  • 5.18.2015

    Got the E.coli strain ET12567 from Professor Li's Lab. Began the experiment about Streptomyces avermitilis.

  • 18th Group meeting

    5.17.2015

    18th Group meeting. Emphasized the lab safety.

  • 5.17.2015

    Did experiments(rainbow kit)related to synthetic biology in Zhejiang Science&Technology Museum with high school students and by this way promoted syn-bio.
  • 5.15.2015

    Read paper and discussed how to enhance the efficiency of Avermectin in Streptomyces avermitilis.

  • 17th Group meeting

    5.14.2015

    17th Group meeting. Discussed the results of the TEM. Tried to contact with a judge from PKU.

  • 5.14.2015

    • TEM scanned our samples: using CNC to pack the Bacillus subtilis.

    • Completed the cover and comic of our proposal which would be handed in to SYNENERGENE.

    • Contacted with Hangzhou Termites control Institute.


  • 5.14.2015

    • TEM scanned our samples: using CNC to pack the Bacillus subtilis.

    • Completed the cover and comic of our proposal which would be handed in to SYNENERGENE.

    • Contacted with Hangzhou Termites control Institute.


  • 5.13.2015

    • Got the genome of tcdA1 and tcdB1 in NCBI.

    • Got TT01(a species of Photorhabdus luminescens) which could produce toxic protein. It would play an important role in our plan B.



  • 5.12.2015

    Successfully checked in termites and tc protein family.

  • 16th Group meeting

    5.10.2015

    16th Group meeting. Communication between dry group and 3 wet groups. Arranged time for experiments.

  • 15th Group meeting

    5.7.2015

    15th Group meeting. Further discussed the proposal and allocated the tasks. Continued to consult professors about several species of bacterium.

  • 5.6.2015

    Gave up synthesizing avermectin in Bacillus subtilis. Tried to overexpress avermectin in E.coli and then transformed its plasmid into Streptomyces avermitilis.

  • 5.6.2015

    Successfully got the solution of CNC!! And it appeared Tyndall effect.

  • 5.4.2015-5.6.2015

    Exams of spring semester.

  • 5.3.2015

    Our termite nest was sent out from Guangdong Province.

  • April
  • 14th Group meeting

    4.29.2015

    14th Group meeting. 10 members would go to Taiwan for NCTU meetup. Results of consulting professors specialized in termites and Bacillus subtilis.

  • 4.28.2015

    • Discussed the plan for embedding baterium with CNC and the self-assembly of CMC and chitosan on the surface of baterium.

    • Debugged the problem of synthesizing avermectin in Streptomyces avermitilis. Consulted professors specialized in synthesizing avermectin in Streptomyces avermitilis.

  • 4.27.2015

    Consulted professors specialized in termites and Bacillus subtilis.

  • 13th Group meeting

    4.26.2015

    13th Group meeting. Discussed something important: the proposal for the 5000€ additional sponsership of SYNENERGENE and visa. A new mamber joined our team. Specified the problem remaining to be solved of each group.

  • 4.25.2015

    Discussd the advantage of Bacillus subtilis campared with E.coli and the utilization of spore of Bacillus subtilis.

  • 12th Group meeting

    4.23.2015

    12th Group meeting. Debugged the previous package plan and decided to pack bacterium with CNC. Planned to buy a nest of termites from Guangdong Province. Discussed several demensions of modeling in this project.



  • 4.22.2015

    Consulted professors specialized in termites.

  • 4.21.2015

    Consulted professors about directly embedding baterium with CNC rather than with the aid of protein domain. Our idea was confirmed and praised.

  • 4.20.2015

    Communication with our instructor. Searched for information about cellulose binding domain (CBDs) and overexpression of ivermectin and avermectin.

  • 11th Group meeting

    4.19.2015

    11th Group meeting. Decided our project: termite terminator. Allocated all members into 4 groups. Each group was responsible for a part of our project.

  • 10th Group meeting

    4.16.2015

    10th Group meeting. Decided to participate in NCTU meetup in summer holiday and apply for the 5000€ additional sponsership of SYNENERGENE. Discussed 3 existing ideas.

  • 4.14.2015

    Preliminarily designed the circuit in Streptomyces avermitilis.

  • 9th Group meeting

    4.12.2015

    9th Group meeting. Kept 3 ideas and the final idea would derive from them. Detailed these 3 ideas from all the aspects.

  • 8th Group meeting

    4.9.2015

    8th Group meeting. Continued to kill unappropriate ideas. Recalled the aim of participating iGEM and reached a consensus.

  • 7th Group meeting

    4.6.2015

    7th Group meeting. Began to kill some undoable ideas and further dug the existing ideas, such as biological enigma machine and killing termites.

  • 6th Group meeting

    4.2.2015

    6th Group meeting. Developed several new ideas, including bio-SCM, biological enigma machine and slow release of certain chemical. Talked about their feasibility.



  • March
  • 5th Group meeting

    3.29.2015

    5th Group meeting. Dug previous ideas and considered their safety and feasibility.

  • 4th Group meeting

    3.26.2015

    4th Group meeting. Continued to share and improve our ideas, such as bio-lens, detection of schistosomiasis and killing termites.

  • 3.25.2015

    Team application completed.

  • 3rd Group meeting

    3.22.2015

    3rd Group meeting. Discussed some ideas about modeling. Shared new ideas.

  • 3.21.2015

    Communicate with Zhejiang Science&Technology Museum.

  • 2nd Group meeting

    3.19.2015

    2nd Group meeting. Built up several rules and principles. Tried to make the most of our BBS.

  • 1st Group meeting

    3.16.2015

    1st Group meeting. Everyone was assigned with his/her own mission. Brainstorm began.

  • 3.15.2015

    Visited the AST space of Zhejiang Science&Technology Museum. Decided to collaborate with.

  • 3.13.2015

    Concluded the winter project.

  • Winter
  • 2.1.2015-3.13.2015

    Everyone read wikis, learned something from previous projects and attempted to come up with new ideas.

  • 1.31.2015

    1st meetup. Had a nice meal~

  • 1.30.2015

    15 members was picked out. ZJU-China 2015 team was set up~ Our story began.


termit