Team:Hong Kong LFC PC/Notebook

PLKLFC_PC

Notebook

Time period Administrative Work Laboratory Work Human Practice
April 2019 Started to have regular meet-up with teammates and teachers every week Discussed which topic was our project’s focus, we had to consider two options: therapeutic approach for brain cancer or for gout. NA NA
May 2019 Settled laboratory duties and positions Checked for any laboratory devices and chemicals which we needed to buy NA Met Professor Chan, who opposed us to investigate brain cancer cells, as it is too difficult to find suitable receptor cell in secondary school level. However, he appreciated the topic of uricase.
June 2019 Read relevant essays to learn more about the mechanism of uricase, uric acid, HIU and variety of plasmids etc. Cultivate two kinds of competent cells, DH5α and BL21. Prepare large amount of agar plates with antibiotics. Met Professor Chui and discuss to learn more details about uricase and decide to adopt intracellular digestion.
July 2019 Ordered materials such as gene sequence and pET-22. However, unlike a journal suggested, pET-22 didn’t ligate with gene of uricase well. Therefore, we order pSB-13 for our experiment after a few weeks. We started our lab work according to our lab workflow, such as PCR, Restriction enzyme digestion, gel electrophoresis, ligation and bacterial transformation Met Shenzhen IGEM Great Bay Team and they encouraged us to use Nissle 1917 instead of yeast as our host cell, since most yeast cannot replicate in the digestive tract. Asked for doctors’ opinions about our project. Worked on a questionnaire about gout to general public.
August 2019 Replenished materials which were used up completely. Ordered new materials for new experiments such as SDS Page, Uric Acid Assay Started to do mathematical modelling. Successfully transformed plasmid with gene of uricase into bacteria. NA
September 2019 Do protein analysis (SDS Page). After discovering some of our genes are not expressed well, we do problem shooting to check whether our SDS page percentage is accurate. We have succeeded this part after modifying the experiment in late September. Analyzed the feedback of questionnaires done in August. Co-organized a workshop with CUHK to share ideas with local secondary and university teams.
October 2019 Finalized and finished mathematical modelling. Completed our wiki page, poster and powerpoint. Ran uric acid assay to prove if ygfu (transporter gene) is present in E.coli. After results of the experiments were finalized, we met Entrepreneur Mr Ricky Chiu for potential business cooperation. Met lawyer Mr Gilbert Chan to check for patent about our project such as uricase. Organized activities such as paying a visit to local elderly centre in order to persuade the general public to accept the probiotic E.coli as a new type of treatment method.