Team:Linkoping Sweden/Notebook

LiU iGEM

Notebook

Notebook

In our Notebook you can follow our progress through the summer and fall of 2018. This notebook contain our weekly reports, for more detailed information and protocols see “Experiments and Protocols”.

Week 24: 11 - 15/6
Summer has begun and we are excited to start our project! This week was filled with preparations for the coming days and weeks. Prepping agar, LB-medium, buffers and so on.
Started working with our biobricks from iGEM followed by methods like transformation in BL21 competent cells, inoculation, plasmid preparation and gel electrophoresis.

Week 25: 18-21/6
This week we continued working with biobricks and changing parameters on our methods to achieve better results. We also had a short vacation as we celebrated the Swedish holiday Midsummer.

Week 26: 15-29/6
This week more investigation on error sources was made as we continued to have no bacterial growth on agar plates. Changing parameters did not have an effect.
We also had a visit from our sponsor Cenova, which told us about their company and also discussed about the IGEM-competition.

Week 27: 2-8/7
This week we begged the iGEM Troubleshooting & Collaborations for help. We got a massive amount of good answers that helped us progress with our project. We also managed to do our first Gibson assembly.
As a part of the iGEM-competition’s criteria; we also started working with the mutagenesis on the plasmid from the previous team (LiU iGEM 2017) and try to improve it.

Week 28: 9-15/7
This week we continued with our Gibson assembly, getting both wanted and unwanted results. This was followed by a lot of changes in PCR temperatures and times. We got good results on our biobrick and chose to verify it on our part pages.

Week 29: 16/7-22/7
This week we learned that extracting DNA from the gel is very difficult if you don’t have good materials to do it. We still struggled with our Gibson assembly but at least our mutagenesis is showing promising results.

Week 30: 23-29/7
This week we really tried to keep a good attitude to the project and to each other as the Gibson assembly kept on failing. Science is a challenge, but a challenge that we love. Our mutagenesis was showing good results as we added restriction sites.

Week 31: 30/7-5/8
We discussed with our supervisor Per Hammarström which led us to the decision to change track in our project. Instead of working with multiple chaperone-systems in the bacterial genome we will try expressing hard-to-fold proteins with just one chaperone, GroES. The chaperone will be induced in E.coli plasmids. We also sent our mutagenesis to GATC Biotech for sequencing, and out modelling-group needed to do some changes with their model, to adjust to the new project.

Week 32: 6/8-12/8
This week we spent a lot of time brainstorming and adjusting our experiments on the new project. While waiting on our new parts from IDT, we worked on our wiki-homepage. We have also recorded a new podcast episode which is a part of our Human practice project - we want to enlighten people about synthetic biology.

Week 33: 13/8-19/8
This week we sent our mutagenesis to GATC Biotech for sequencing and the new part from IDT finally arrived and we are therefore back in the lab. We also colored our “Liu-IGEM” logo on campus, which is a tradition for students unions at Linköping University.

Week 34: 20/8-26/8
This week we spent a lot of time in the lab and did digestions and ligations of GroES and also heat shocked our mutagenesis.

Week 35: 27/8 - 2/9
This week we struggled to get our results, with no good results in sight…

Week 36: 3/9-9/9
For some of us in the team, a new semester has begun and we have to study and try to work with iGEM continuously. We finally see some progress on our results and continue working with GroES and our mutagenesis.
This week we have attended the annual reception called “Kalas” which is an event for new students at Linköping University to get attached with different student societies. On this event, we told students about our team and about the competition and recruit new members to our student organisation.

Week 37: 10/9- 16/10
This week, the LiU-iGEM team went to central Linköping to find participants for our survey and enlighten people about synthetic biology and about our project.

Week 38: 15/9 - 21/9
We are trying to get our results we want to achieve our goal before the Wiki-freeze . We are still doing some mutagenesis and working with a collaboration with Lund which is about exchanging our Biobricks and try to co-express them. We are doing some SDS-page which takes forever to do, but we are hoping for good results.

Week 39: 22/9 - 28/9
This week we sent in our Team banner, which is something the teams are going to see in the Giant Jamboree in Boston. (We hope you will listen to our presentation on thursday 25/10 and come to our poster to discuss our project!).

Week 40: 29/9 - 7/10
This week our team member Martin Viksten had a TEDx-talk in Norrköping about iGEM and about our vision. In the lab we had progress and sent our mutagenesis to Germany for sequencing. We also started to do the protein-expression of our co-induction system. The system utilizes chaperone expression to increase protein folding. The proteins we co-induced with chaperones were aggregation prone proteins. We also continued doing our wet-lab collaboration with Lund.