Period 1: 2023.1.10-2023.4.30:
Because we come from different areas in China, we are planning to learn some knowledge online together based on our teachers and development of project plan. We have learnt as below:
1) the iGEM Project manual
2) Books on Scientific Research and synthetic Biology
3) Concept, technical vocabulary, text and video materials
4) The charm of synthetic biology
5) In-depth Analysis of iGEM Award-winning cases
6) Thinking and skills of scientific research
7) Introduction to iGEM Competition
8) Introduction to Synthetic Biology
9) Introduction to Molecular Biology
10) Introduction to Cell Biology
11) Skill Training: Literature Retrieval Methods and Skills
12) Skill Training: Human Practices Method
13) Experimental Methods and Skills Learning in Synthetic Biology
14) Skill Training: Modeling and Data Processing
15) iGEM Awards Criteria Analysis (Entrepreneurship/Human Practices）
16) iGEM Awards Criteria Analysis (Life science)
Period 2: 2023.7.25-2023.8.15:
We begin to do the tasks separately. For wet laboratory activities, we had prepared experimental research & divide tasks、conduct experimental research & record data with mentors team、conduct literature research & compare experimental methods、conduct experiment records & develop professional lab habits、analyze experiment result & process、work with dry team member to conduct promotion & education work etc.
Some details are below:
Stage1:
1) Familiar with laboratory equipment and drugs and learn about laboratory safety matters；2）Introduction to laboratory instruments and drugs；3）Laboratory safety precautions；4）Configure required reagents；5）Organize supplies；6）Prepare culture medium.
Stage2:
1）Competent cell preparation and recombination and Principles of plasmid transformation；2）HIV-1 RRE plasmid transformation；3）Receptive cell transformation；4）Coating plate culture.
Stage3:
1）Obtaining the HIV-1 RRE gene sequence；2）RNA gene analysis processes and methods；3）Explain the acquisition of HIV-1 RRE gene sequence；4）Explain the RNA gene analysis process and methods.
Stage4:
1）Single bead selection culture；2）Shake the bacteria and take samples regularly to measure the growth curve (OD value)；3）Regular sampling to measure growth curve (OD value)；4）Seamless cloning technology explained；5）Purify plasmid and measure concentration.
Stage5:
1）Primer design principles and precautions；2）Nucleic acid electrophoresis and photography；3）Analyze electrophoresis results；4）HIV-1 Rev protein plasmid preparation)；5）Competent intracellular transformation；6）Plate overnight culture.
Stage6:
1） Observe the growth of strains on the plate; 2）Pick a single strain and shake it for culture；3）Take samples regularly to measure OD；4）Preparation of DNA template for PCR amplification reaction；5）Nucleic acid electrophoresis verification template, photos
；6）Preparation of HIV-1 Rev Glycerol Bacteria Preservation；7）Pick a single colony again and culture it overnight to prepare for plasmid extraction the next day.
Stage7:
1）T7 RNA polymerase in vitro transcription principle；2）Transcription reagent preparation and configuration；3）HIV-1 RNA in vitro transcription；4）Purification of in vitro transcribed RNA；5）RNA electrophoresis, photography；6）HIV-1 Rev shaken, overnight.
Stage8:
1） HIV-1 Rev protein expression；2）Regular OD measurement；3）Regular OD measurement again; 4）Measure OD regularly to 0.6-0.7, IPTG induces expression; 5）Bacteria collection.
Stage9:
1）Break bacteria, extract Rev protein, take samples, and retain samples to verify Rev protein expression；2）Configure protein purification gradient buffer；3）Protein column purification；4）Collect the purified fractions of the target protein Rev and verify by SDS-PAGE protein electrophoresis.
Stage10:
1）Experiments on RRE and Rev protein interactions at different ratios；2）EMSA conducts interaction verification; 3）Electrophoresis results and photos.
Stage11:
1）Explain the acquisition of protein gene sequences；2） Compare the analysis processes, functions and methods of protein and nucleic acid RNA；3）Selection and research significance of RNA and protein action sites.
Period 3: 2023.8.10-2023.9.20:
For dry laboratory activities,we had conducted social research& field research、conduct expert Interview、establish team brand promotion channel & promotion、fundraising & business entrprenuership、outreach & collaboration/communication、film team promotion videos & edit etc.
Stage1:
1）Familiar with laboratory equipment and drugs；2）Learn about laboratory safety matters；3）Team name, logo design；4）And peripheral and product promotion product design；5）Explanation of experimental principles；6）Laboratory tour；7）Laboratory safety precautions；8）Improve team culture product content
Stage2:
1）Create team IP；2）Complete peripheral order purchases (financial statement records), etc. 3）Conduct social research on the topic, including questionnaire design and preparation for social interviews；4）Formulate team work plan (publicity, science popularization, education, charity； 5）Completed social research, collected feedback data, and completed the first content release on publicity channels.
Stage3:
1）Design and planning of education and publicity activities (which groups are targeted, educational activity methods, required materials); 2）design and planning of Inclusive Award activities；3）Dry and wet team communication; 4）Bio-bank visit; 5）Optimize education and publicity activity plans.
Stage4:
1）Project solution acceptance questionnaire design, release preparation, expert interview questions；2）The first expert interview (technical feasibility) and feedback recording; 3） the completion of the second publicity channel content release.
Stage5:
1）(Collaboration) Contact other iGEM teams to confirm joint activity plans；2）Complete the design and preparation of promotional posters, leaflets and other materials；3）Communication and discussion on the progress of wet and dry teams；4）Team promotional video idea.
Stage6:
1）Outbound implementation of education and publicity activities；2）Activity summary；3）Writing the wiki: Communication section；4）Write a team video script；5）Completed the third promotion channel content release.
Stage7:
1）Promotional video material shooting；2）iGEM team cooperation and communication；3）Business planning explanation；4）Project business concept and product design.
Stage8:
1）Plan fundraising events；2）Prepare questions for second expert interview (product application)；3）Second expert interview and feedback record；4）Optimize commercial product design；5）Completed the fourth promotion channel content release.
Stage9:
1）Write a business plan；2）Wet and dry team communication；3）Team Booth Poster Design；4）Wiki web interface concept design.
Stage10:
1）Team promotional video shooting and editing；2）Conduct fund-raising planning activities and sustainable development creative activities；3）Completed the fifth promotion channel content release；4）Write a business plan；5）Impact part of the content.
Stage11:
1）Check for omissions and fill them in, and prepare relevant materials；2）Clarify tasks and plans for subsequent online activities.
Period 4: 2023.9.10-2023.10.10:
Building the Team Wiki. Our team members begin to record the team wiki based on our performances we have done before.
The experiments we have used as below:
1）Molecular experiments
2）Explanation of seamless cloning technology (including primer design, etc.)
3）DNA extraction and testing
4）PCR amplification of target genes
5）PCR product recovery-vector ligation-competent cell transformation
6）DNA/RNA electrophoresis
7）RNA in vitro transcription
8）Obtaining gene sequences and analyzing processes and methods of gene functions
9）Screening and significance of RNA-protein interaction sites