Team:NTHU Formosa/Collaborations

Collaboration with NTHU_Taiwan

Collaboration with NTHU_Taiwan

  As a team composed of members from biology background, we gladly offer help and troubleshoot the problem on making constructs for team NTHU_Taiwan, being insufficient with members from biology department. Since August 3rd, we had several meetings with 2018 iGEM team NTHU_Taiwan.

  Earlier before the meeting, NTHU_Taiwan designed the gene of insert for their experiment. After getting the synthesized gene part from IDT and digesting both the inserts and the vectors, they proceeded to ligation, which failed after times they tried.

  We went through details from their design of the gene, cutting site and protocols. Later we concluded that the major reason for failing to make the constructs is due to low concentration of the inserts left following digestion and purification.

  We re-designed primers for NTHU_Taiwan and suggested NEBuilder Assembly kit, so that they can avoid the loss of their inserts while digesting and purifying the DNA. The kit requires lower DNA input and can save time by avoiding PCR amplification steps.

  A few weeks later, we were informed that NTHU_Taiwan still failed to make the constructs. Therefore, we provided our lab for their experiments and kept an eye on them through out their experiments to make sure that there’s no man-made technical false.

 Gladly, after several meetup with us and our assistance in lab, team NTHU_Taiwan successfully made all three constructs they needed-- pSB1C3 - AHL snesor - Anti – STAR, pSB1C3 - pLac – STAR, pSB1C3 - pLux – GFP. It was very pleasant experience to work with NTHU_Taiwan.

  Since we urge to seek help in building wiki page, in return, NTHU_Taiwan also give us host a meeting with us teaching coding and giving us tips that help making wiki page easier for us. We took the wiki pages designed by previous teams as examples to learn and practiced basic coding skills(Fig. 1).

Fig. 1. NTHU_Taiwan helped us making wiki page.

Fig. 2 & 3. NTHU_Taiwan validating construct used in our experiment.

Fig. 4. Constructs from the left to right: pCAG-GBP, pTRE3G-LuxAB, pCMV-LuxCDEfrp and pTRE3G-mCherry. The size of the bands is correct.

Collaboration with NCTU_Formosa

Collaboration with NCTU_Formosa

 Since we are a fairly new iGEM team, we are not familiar with the details about iGEM competition. Therefore we turned to NCTU_Formosa, who hosted this meetup on Aug 24th, during which we took turns presented our project, gave each other feedbacks and suggestions. NCTU_Formosa is especially very helpful on our wiki page. As almost all of our team members are in biology background, building wiki page is truly a huge challenge for us. In this meeting, NCTU_Formosa spent a lot of time teaching and troubleshooting our problem on building wiki pages (Fig. 1), for which they prepared a 2-hour-course containing content list below. This is truly a very successful and helpful collaboration. After the meetup, NCTU_Formosa stayed in contac with us by messenger to further assist us with constructing wiki page.

Team NCTU_Formosa gave us a tutorial about html. During the tutorial, we learned:

1. Basic tags and elements for html
2. Image scale and website format adjustment
3. Styling html with CSS and JavaScript
4. Conversion of websites in PC and mobile version

Fig. 1. NCTU_Formosa taught us building wiki pages.

Thanks to NCTU­_Formosa’s guide, discussion and their generosity to assist our problems through messages after the meetup. We both learned a lot from each other.

Fig. 2. Members of NCTU_Formosa and NTHU_Formosa.

 Since NCTU_Formosa is very interested in our project and we both would like to enhance our collaboration, we validated each other’s parts using in the experiments. NCTU-Formosa validated the DNA constructs we expressed in 293T cells in our experiments (Fig. 3).

Fig. 3. NCTU_Formosa running gel electrophoresis for us to validate our constructs.

Constructs from the left to right: pCAG-GBP, pTRE3G-LuxAB, pCMV-LuxCDEfrp and pTRE3G-mCherry. The size of the bands is correct(Fig.4).

Fig. 4. The size of the bands is correct for all samples using digestion check.

We also validated the protein, bacteriocin, for NCTU_Formosa(Fig. 5).
M: marker
C: control
E1: enterocin B (35.5kDa)
E2: enterocin 96 (35.9kDa)
Enterocin B and Enterocin 96 are bacteriocin produced by bacteria that can regulate the growth of specific strain. The results showed that both protein are correct(Fig. 6).

Fig. 5. We ran the SDS-PAGE for NCTU_Formosa to validate their protein.

Fig. 6. The result of the samples are correct.