After our first group meeting, we were divided into 4 groups and got the first assignment since we join the team — read as much wikis as we can and came up with embryonic form of the new project during the winter holiday.
From 21st to 26th, our team members held a meeting to report the wikis we have read before and to communicate our ideas about the project. We also learned some skills that we should have in the laboratory.
Raised plans for future work to get sponsorship.
Ran our official account on Wechat and published the first article on 31st January .
In this month, we were going to express hBD3 in the periplasmic space of E. coli at first, but after a period of study, we think that’s an unfeasible way.
On April 22, we went to the Laboratory of Molecular Biology, Shenzhen PKU-HKUST Medical Center and took some experimental training. On April 25, we met Dr. Haoqian Zhang, a professor in Chinese Academy of Sciences, and discussed about our
project. He gave us much advice.
Being inspired by the instructors, we found some serious problems that may affect our future study, for example: 1) How to avoid the formation of inclusion bodies? 2) How can we make sure the molecular chaperones service can work as expected?
3) The feasibility of the project is actually low. What can we do?
There were also many problems found in Human Practice, like: 1) How to dispel the discrimination of that disease in public? 2) How to make the medicine more convenient for people to use? 3) How to shorter the time for treatment?
On April 1st, we released a questionnaire about Trichophyton rubrum.
On April 20th, we made a wonderful speech about our project and synthetic biology at Scientific Festival in Shenzhen Foreign Languages School.
Opened an account on Twitter and then published our articles on both Wechat and Twitter.
On April 29th, we met up with universities team and high school teams from south eastern China at Shenzhen University.
On April 30th, we closed the website of Trichophyton rubrum questionnaire.
After the human practice part, we had a profound discussion about changing the project. Finally, we changed our project into the treatment for cancer in this month.
On July 5, we designed our Single-Guide RNA (sgRNA) which could specifically bind to the mutant BRAF gene.
On July 8, we started to culture the melanoma cell lines A375 and G361.
On July 9, we cultured the bacteria containing our plasmid containing our sgRNA and Cas9 protein.
On July 11, we extracted genome from A375 and G361 cell lines to check if the cell lines had BRAF V600E mutation. We extracted the plasmids to prepare for cell transfection.
On July 13, we transfected the cells with our plasmid.
On July 14, we assigned the task of writing different parts of our wiki to everyone and started writing our abstract, description and design of our wiki.
We cultured the cell lines.
On July 20-21, we submitted 4 parts.
On July 26, we cultured the cells in a 6-well plate and transfected the cells in order to measure cell migration by wound healing assay.
On July 27, we seeded the cells in a 96-well plate and trasnfected the cells in order to measure the cell proliferation by Transdetect Cell Counting Kit (CCK). We planned to scratched wounds across the cells but failed, for G361 cells didn’t
cover the dish.
On July 28-30, we stained the cells to detect the results of CCK assay. We cultured the cells.
On July 31, we finished writing our first draft of our abstract, description and design.
Attended the 19th International Botanical Congress.
On July 10th, we met up with Southern University of Science and Technology iGEM team.
From July 24th to July 30th, we went to Bali island to teach teenager “How to keep healthy”.
On July 30th, we delivered a public speech about iGEM, Synthetic Biology and Melanoma in Shenzhen Bookcity and gave out questionnaires while the speech was going on.
On Aug 1, we exchanged the Cell medium. We froze the Cells. We seeded the cells in a 6-well plate and transfected the cells to measure the cell migration by Transwell assay. We seeded the cells in a 96-well plate and transfected the cells
in order to measure the cell proliferation.
On Aug 2, we stained the cells to detect the results of CCK assay. We stained the cells to detect the results of Transwell assay.
On Aug 3, we subcultured the melanoma cell, exchanged the cell medium and stained the cells to detect the results of CCK assay.
On Aug 4 – 5, we stained the cells to detect the results of CCK assay and subcultured the melanoma cells.
On Aug 8, we seeded the cells in a 6-well plate and transfected the cells to measure the cell migration. We cultured the bacteria containing our plasmid for plasmid amplification.
On Aug 9, we extracted the genomic DNA of the cells. We transfected the cells.
On Aug 10-12, we seeded the cells in a 6-well plate and transfected the cells for TA cloning assay.
On Aug13, we seeded the cells in two 6-well plate and transfected the cells for cell apoptosis and migration assay.
On Aug 14, we transfect the cells, cultured the bacteria containing our plasmid for amplification, stained the cells to detect the results of CCK assay and flow cytometry assay.
On Aug.15, we extracted the plasmids and stained the cells to detect the results of Transwell assay.
On Aug19, we attended the meet up and introduced our project.
In the whole August, we gained a lot from experiments.
From Aug 19th to Aug 20th, we cooperated with Shenzhen College of International Education to organize a meet up with several iGEM teams.
Completed the Attribution part.