Team:JNFLS/Notebook
JNFLS
Notebook
Week 1(4.9-4.15)
★ Recruitment of team members and first meeting.
★ Brainstorming and learning about relevant materials.
★ We started to discuss our ideas and invited teachers to give some advice about our projects.
★ Presentation within the team to share what we learn about synthetic biology.
Week2(4.23-4.29)
★ Finally,we choose the idea of curing colon cancer based on the technique of Crispr/Cas9 came into being.
★ A lab was provided by Shandong University in summer vacation.
★ After many discussions, everyone's division of labor is becoming clearer.
★ We began to use our spare time to study experimental techniques.
★ We started to brainstorm in order to create our team logo.
Week3 (5.13-5.20)
★ We had a safety education before we started the formal experiment, including lab safety, researcher safety and environment safety. And we learned some emergency responses, such as how to use extinguisher and fire blanket, and how to use emergency shower and eye wash.
★ Made a concrete plan for our project.
Week4 (5.21-5.27)
★ Extracting of pcDNA3.1.
★ Preparing theTAT plasmid
★ Learning cell culture techniques, and beginning to culture of colon cancer LOVO.
Week5 (5.28-6.3)
★ The construction of plasmid vector pcDNA3.1-TAT.
★ We have decided to build our parts and primers sequence was designed.
★ CRISPR plasmid was obtained, and after transformation, monoclonal selected, we have sent plasmid to the company for sequencing verification.
Week6 (6.4-6.10)
★ After connecting the cleavage products, we have sent them to the company for sequencing.
★ We started building two parts using PCR technique and digestion technique
★ According to our design, We should know the sensitivity of the tetr repressible promoter. So we decided to build two plasmids.
Week7 (6.11-6.17)
★ Successfully constructed and cryopreserved.
★ Continuing to build part: 1A and 1B.
★ We went to Hebei to have teaching activities for a week to spread IGEM to children in countryside.
★ Preparing of experimental materials.
Week8 (6.18-6.24)
★ Learning cell culture techniques, and beginning to culture of 293T cells.
★ According to our topic, we designed a series of questionnaires.
★ We began to construct the first plasmid vector
Week9 (6.25-7.1)
★ It was verified by sequencing that we have building parts 1A and 1B successfully.
★ Through a small range of issuing questionnaires, we have further modified our questionnaires.
★ Continuing to construct the first plasmid vector, and preparing the materials for the second plasmid vectors.
Week10 (7.2-7.8)
★ We went to Qi Lu Hospital to have a meeting with the doctors who specialized in colon cancer cuing.
★ Extracting of luciferase and began to monitoring data.
★ We started building two parts using PCR technique and digestion technique
★ The construction of plasmid vector was started.
★ We started issuing questionnaires in a wide range.
Week11 (7.9-7.15)
★ Continuing to Construct the plasmid vector.
★ Preparing for the work of interlab.
★ It was verified by sequencing that we have constructed the first vector successfully. And we began to constructed the second vector: ROO40+E0840.
Week12 (7.16-7.22)
★ Continuing to build part: 1C,1Dand 1E.
★ Continuing to Construct the plasmid vector.
★ For the interlab, we have carried out preliminary experiments to be familiar with the use of instruments.
★ Continuing to construct the first plasmid vector.
Week13 (7.23-7.29)
★ Extracting of luciferase and beginning to monitor data.
★ It was verified by sequencing that we have building parts 1C,1D and 1E successfully.
★ We have done the work of interlab.
★ we added a series of tetracycline with a concentration gradient to some of the tubes with the same initial fluorescence intensity and measured the change in its fluorescence intensity.
Week14 (7.30-8.5)
★We have interview clinicians to know more about cancer and make some adjustments on our experimental ideas. ★The plasmid vector was finally constructed successfully after hard working, and we decided to transfect the plasmid into the cells. ★It was verified by sequencing that we have constructed the secong vector successfully. Week15 (8.6-8.12) ★We have went to Qingdao to communication thoughts with the students of Chinese Marine University. ★We have made a series of posters. ★After data processing, we got the curve which showed the influence of the tetracycline on the fluorescence intensity of TetR fusion protein. Week16 (8.13-8.19) ★Continuing to build part: 1F and 1G. ★Learning cell transfection technology. ★Public propaganda of science in the Technology Museum. ★We transformed these two plasmids to competent cells, and according to their different resistances, we selected the strain of interest. Week17 (8.20-8.26) ★It was verified by sequencing that we have building parts 1F and 1G successfully. ★We have participated the CCIC and acquired much valuable experience. ★We went to science museum to spread the idea of iGEM and hand out questionnaires. Week18 (8.27-9.2) ★We take part in the CCiC in Fuzhou and acquired valuable advice. ★Extraction of luciferase and began to monitoring data. ★We started building two parts using PCR technique ★Extracting the genomic DNA from cells, and starting exploring the best conditions of PCR. ★We added a series of tetracycline to combined with TetR in order to relieve the inhibition of TetR on TetR repressible promoter. Week19 (9.3-9.9) ★We have launched a series of activities to promote the IGEM competition for Freshmen. ★A presentation for the students in our own school ★ Week20 (9.10-9.16) ★Completing the data determination of cell NHFB. ★Continuing to build part: 1H and 2A. ★The PCR products were identified by restriction enzyme digestion. Unluckily, we didn't get the expected results. After analyzing the whole process, we have changed some conditions and decided to do it again. ★We started writing wiki. ★we started measuring the fluorescence intensity of cells at different tetracycline concentrations over time. Week21 (9.17-9.23) ★It was verified by sequencing that we have building parts 1H and 2A successfully. ★Six team members were select to join the Giant Jamboree. ★continuing to measur the fluorescence intensity Week22 (9.24-9.30) ★Starting to analysis the data we have monitored, and looking for help in modeling. ★We have changed the annealing conditions to separated the double chains. ★The collection of the data from questionnaires, and summarizing the main trend. Week23 (10.1-10.7) ★Sorting out experimental ideas and establishing correlation model. ★We have processed the data of questionnaire survey. ★After many measurements and data processing, we have realized the sensitivity of TetR repressible promoter. Week24 (10.8-10.14) ★Continuing to build part: 2B. ★We changed the reaction time of the enzyme digestion. ★We have designed our winter uniforms. Week25 (10.15-10.21) ★The experiment was almost end and the relationship between starting efficiency of hTERT and survivin promoter in different cell lines and time after transfection was Investigated. ★It was verified by sequencing that we have building parts 2B successfully. And today we have sent all of our parts to the Genscript, Nanjing, China.