Team:KU Leuven/Notebook
Notebook
I know what you did last summer! KU Leuven iGEM Team 2017 has gone through some ups and downs, but that didn't stop them from being creative and to explore the world of science. Other than that, a lot of fun activities were planned. To know more about their adventure, feel free to go down the memory lane with us.
21 - 27 Feb
First Meeting
This is the meeting where the team spirit was established and missing pieces of our adventure to success were collected. For example, who is going to be our PI? How do we get sponsors? Who will be our leader to guide us all the way through to the end of the project? Every team member was assigned a subcategory, which they will complete with passion and dedication. A very subtle move towards possible project ideas was made during this meeting.27 Feb - 7 Mar
Project Ideas
This week, we discussed the many proposals about project ideas along with some professors. There were some remarkable ideas on the table: Development of an ecosystem in a space station, DNA fitting to predict an individual’s health, weight and nutrition, introduction of a filter over intestinal villus for better uptake of Gluten in Coeliac patients, and more.
We also completed our team registration and team leaders were chosen for the subcategories. Nothing could stop us from entering this!7 - 13 Mar
Brainstorming
The voting deadline for project ideas was nearing. In the meeting, a lot brainstorming happened and a lot of new ideas were added to the list. The layout for a sponsor brochure was also created. We reminisced about our childhood and our first contact with science for inspiration for how we might educate the local children. During the meeting, one of the students who is an avid traveler booked flight tickets.
Boston, HERE WE COME!
14 - 21 Mar
A visit from two familiar figures
This week, we had a very special meeting. Two ex-iGEM members came over and gave us advice about their struggles and experiences in their iGEM team last year. A lot of stories were shared and the trust of KU Leuven was handed over to the current iGEM team as every member got an account and joined the KU_Leuven team on igem.org. There is also a rumor going around, which tells us that six professors are going to visit us within two weeks!
21 - 27 Mar
Sponsor Brochure
The first activity of the iGEM Team 2017 was planned, which is having dinner together in the following week. An electrophysiology professor came to visit the KUL iGEM Team, which is a great honor. A lot of ideas were exchanged and it gave us a better insight into our potential project. The requirements for multiple projects are being checked and the final touches to the sponsor brochure was done. Sponsors were reaching out to the KUL iGEM Team and mutually beneficial agreements have been made.28 Mar - 17 Apr
3 Projects and 3 Teams
This week, which is also the last week before spring break, the team was divided into three groups, each working on a potential project. Multiple methods and perspectives were discussed, pushing our creativity to the limit. We planned two meetings with potential sponsors which wrapped up the first part of preparations before spring break hit in.
18 Apr
Arrival of the DNA kit
While it is still half a year apart from the Giant Jamboree, the team was already been looking for accommodations in Boston. Visas were applied for, rooms were booked and our summer schedule slowly materialized much like the sunshine in Belgium. As our final chosen project slowly got its shape, students explored their inner creativity and set up a potential draft for their logo. Our DNA distribution kit from iGEM has arrived and we were so excited!29 Apr
KUL iGEM goes VIB
The KULeuven iGEM Team 2017 got the opportunity to spread a word about iGEM on the VIB conference with over 700 attendees! Two courageous teammates did a presentation and it was a nerve-wracking day for them. The KULeuven Team was there to support them and they nailed it! Congratulations and thank you to Pieter-Jan and Jana! 10 - 26 June
iGEM vs Exams
Despite the fact that the students are in the midst of the exams, impromptu meetings were planned to iron out the last details about the project. We set the summer schedule and chose a logo. We even squeezed in some fascinating scientific articles during our exam study breaks.
27 June
KUL iGEM goes Wageningen
The KULeuven iGEM team got an invitation from the iGEM Wageningen team for the iGEM Benelux meet up! Teams from Groningen, Delft and Louvain joined in and the KU Leuven Team attended many interesting presentations and even went home with a package of knowledge, inspiration and new ideas. Thank you to Wageningen for having us and the team from Groningen for inspiring us with your 3D phage model!28 June
Start of the Labwork
All the members of the KU Leuven iGEM Team 2017 have received a short introduction about lab safety.29 June
Stable mHCN-HEK cell line defreeze and introduction to HEK cell splitting
The iGEM summer has finally started! The students collected information and shared them with the teammates. Remarkable scientific articles have been discussed and the very first experiments were carried out. Two cell lines, stable HCN HEK cells and hERG cells were established and transfected into four categories, as follows:
Fura-2-AM was added and the cells were visualised with Ca2+ imaging. We obtained immediate results and discovered significant differences in cells that were transfected with both α1G and hERG. Oscillations with a duration of 12 seconds were observed in the cells. Following the advice from an electrophysiology professor, we managed to optimize their transfection and create a better transfection efficiency and ratio by stimulating HCN channels with cAMP, causing more and faster oscillations and speeding up the process by giving stronger signals. Measurements were improved and made faster by using only 1 wavelength.30 June
Splitting and transformation
The team members were taught in how to split HEK cells.
On the 3rd of July, the team members has created their first primer design and from 4 - 5 July, cells were transformed with the following plasmids:
• pCNDA5/FRT/hErg
• pCAGGSM2/IRES/GFP/ShIR
• pCNDA5/FRT/mHCN
• pCAGGSM2/IRES/GFP/Nav1.5
• pCAGGSM2/IRES/GFP/a1G
• pCAGGSM2/IRES/GFP/a1C
• pIresRed
On 6 July, a midiprep was done for the purification of the DNA. Then HEK cells were transfected for Ca2+ imaging.7 July
Ca2+ imaging
mHCN and hERG cell lines are being practiced on with Ca2+ imaging.10 July
Koffiekoeken!
As we all know, Monday is the beginning of a great week, but also one of the hardest mornings to get out of bed. Luckily, Liesbeth, one of the teammates, is like a ray of sunshine. That morning she provided the team with fresh and tasty pastries, along with a pretty smile.11 July
First Patch Clamp training
A few students from the team are learning the Patch Clamping technique. Their curiosity and excitement was written all over their faces as they discovered ion channels on their first day in the electrophysiology lab.They were being trained in Patch Clamping with TRPM3 cells and it took from 11 - 26 July.12 July
Unboxing
Pieter-Jan, one of the students of the team, was completed buried under the boxes. We were stocked with a variety of equipment, thanks to our sponsors.20 - 26 July
hERG, mHCN and SHIR
On the 20th, agarose gels were being prepared and PCR samples were being loaded on the agarose gels, but no results were seen.
On the 21-24th, PCR was carried out with the cloning protocol for hERG, mHCN and SHIR, and results has been achieved for hERG.
On 25 July, the PCR product with hERG was purified.
On 26 July, restriction digestion of hERG and its plasmid backbone was carried out.
On 27 July, HEK cells were transfected with: AIG, mHCN and hERG.28 July
Ligation hERG/DSRed Vector
Start of ligation with hERG and DSRed Vector. First time patching with portable patch on our hERG cells that are transfected with AIG and mHCN. Calcium imaging was used to test stable HCN and hERG cells with different potassium concentration.
On 1 August, hERG was transformed.Date
Transformation
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