This Notebook page will help you understand what we have suffered done in these days.
May
May
Week01: 05/03-05/09
WL:
Explore the mechanism how TAT and penetratin function
Explore the mechanism how S. aureus resist antimicrobial peptides
Find out whether antimicrobial peptide harm the organelles
Confirm the latency of S.aureus in professional phagocytes
DL:
Determine the materials in each dressing layer
Integrate the preparation of dressing, such as protocol, equipment, etc.
Design the thrombin cleavage assay
HP:
Complete the parent-child picture book’s outline
Prepare the teaching plan for high school and elementary school students
Operate our Instagram and Facebook
Seek for collaboration opportunities among iGEMers
Write a promotion video script
Start planning a fundraising plan
Week02: 05/10-05/16
WL:
Search the lysosomal enzyme candidates for releasing intracellular AMPs
Cloning eGFP-TAT and eGFP into expression vectors-PCR, TA, and sequencing confirmation
DL:
Order the material of dressing
Confirm the connection of each layer of dressing
Design the thrombin cleavage assay
HP:
Rehearsal the experiment in the high school teaching plan
Shoot promotion video
Start to make our parent-child picture book
Prepare for interviews with domestic experts
Design the flyer for the teaching plans
Week03: 05/17-05/23
WL:
Design the construct of biobrick
Search the the activity of AMP in different pH level
Search the cleavage preference of lysosomal legumain
Cloning eGFP-TAT and eGFP into expression vectors- ligation into expression vectors
DL:
Determine the protocol and equipment of thrombin cleavage assay
Thrombin cleavage site prediction
Determine absorptive layer and collagen layer protocol
Determine the container and size of the dressing
HP:
Put our teaching plan on record, responding to the epidemic
Week04: 05/24-05/30
WL:
Explore how antimicrobial peptide functions
Explore the cleavage preference of lysosomal enzymes-cathepsin H, F, D, S, L
Explore the biosafety of cell penetrating peptides
Cloning eGFP-TAT and eGFP into expression vectors- ligation into expression vectors
DL:
Design the thrombin cleavage assay
Preparation of absorptive layer
HP:
Make PowerPoint for the teaching plan
Finalize the parent-child picture book
Design our team logo
Design team’s peripheral product
Design postcard(the cooperation with iGEM-Duesseldorf)
Start to cooperate with the other iGEMer for translating our parent-child picture book
June
June
Week05: 05/31-06/06
WL:
Explore how antimicrobial peptide functions
Find the the macrophage culture and stimulation protocol
Explore the biosafety of cell penetrating peptide and antimicrobial peptide
Find evidence supporting the severity of MRSA latency
Write the safety form.
DL:
Design the thrombin cleavage assay and confirm protocol and equipment
Confirm and modify the preparation principle of absorptive layer
HP:
Online meeting with iGEM Ioannina
Prepare for the parent-child online picture books
Design team uniform
Week06: 06/07-06/13
WL:
Search the mechanism of antimicrobial peptide-finished
Find the protocol of hemolysis
Explore the biosafety of cell penetrating peptide and antimicrobial peptide
Find evidence supporting the severity of MRSA latency
Write safety form
Ligate the eGFP-TAT to pET15b
Perform the PCR for CBD-linker, thrombin cleavage site for AMP/TAT mimic release, and thrombin releasable AMP
Perform the colony PCR of TA-CBD-linker, TA-Thrombin cleavage site for AMP/TAT mimic release, TA-thrombin releasable AMP
DL:
Confirm the human thrombin concentration
Preparation of absorptive layer
Order collagen solution
HP:
Trim the promotion video
Draw a comic about our project(the cooperation with iGEM IISER Berhampur)
Week07: 06/14-06/20
WL:
Explore the biosafety of cell penetrating peptide and antimicrobial peptide
Find evidence supporting the severity of MRSA latency
Write safety form
Perform the colony PCR for TA-CBD-linker, TA-Thrombin cleavage site for AMP/TAT mimic release-eGFP, TA-thrombin releasable AMP
Perform the digestion for TA-CBD-linker, TA-Thrombin cleavage site for AMP/TAT mimic release-eGFP, TA-thrombin releasable AMP
DL:
Prepare different mixing ratios of the absorptive layer to find the suitable group
Wiki team member page design
HP:
Prepare a collaboration proposal for Taiwan iGEM Conference
Dub the promotion video
Invite Hsin-Yi Foundation as our interviewee
Hsin-Yi Foundation regards the promotion of early childhood education and parenting education as their mission
Week08: 06/21-06/27
WL:
Fighting with midterm examination!!!!!!! XD
DL:
Fighting with midterm examination!!!!!!! XD
HP:
Prepare for a meeting with iGEM Team KOR_HS
Contact Adien Lo, a 17 years old podcaster, to see if we can collaborate
Send out postcards to Germany for worldwide iGEMer’s postcard exchange activity launched by iGEM Team Duesseldorf
July
July
Week09: 06/28-07/04
WL:
Explore the cleavage preference of lysosomal enzymes-cathepsin B, D.
Explore the biosafety of cell penetrating peptide
Find evidence supporting the severity of MRSA latency
Cloning eGFP-TAT and eGFP into expression vectors- ligation into expression vectors
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP, and thrombin releasable AMP into expression vector- ligation into expression vectors
DL:
Find the suitable mixing ratio of alginate and chitosan solution to prepare our absorptive layer
Confirm Swelling test protocol
Confirm thrombin unit conversion
Wiki team member page and homepage design
HP:
Finish the parent-child picture E-book, and start to translate it into an English version
Participate in Taiwan iGEM Conference hosted by iGEM Team CSMU_Taiwan
Communicate with Kinmen Hospital to seek professional advice about infection data
Invite Junyi Academy as our interviewee
Junyi Academy creates an online learning platform for students from elementary school to college
Week10: 07/05-07/11
WL:
Explore the cleavage preference of lysosomal enzymes-cathepsin B, D
Explore the biosafety of cell penetrating peptide
Find evidence supporting the severity of MRSA latency
Cloning eGFP-TAT and eGFP into expression vectors- ligation into expression vectors
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP, and thrombin releasable AMP into expression vector- ligation into expression vectors
DL:
Prepare absorptive layer by different concentrations of alginate and chitosan solution
Conduct the swelling test to compare water uptake ability of the absorptive layer
Collagen layer protocol
Convert the unit of commercial thrombin to human thrombin and confirm its usage in our experiment
Order thrombin
Design Wiki style
HP:
Have an online meeting with the iGEM Team KOR_HS
Dub the online teaching video
Do the market research of our parent-child picture book
Turn our parent-child picture E-book into the paper one
Finish the promotion video
Translate parent-child picture book into a Cantonese version, thanks to iGEM Team HKUST
Week11: 07/12-07/18
WL:
Explore the mechanism of antibiotic-resistance of S. aureus
Explore the biosafety of cell penetrating peptide
Organize evidence supporting the severity of MRSA latency
Cloning eGFP-TAT and eGFP into expression vectors- ligation into expression vectors
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP, and thrombin releasable AMP into expression vector- ligation into expression vectors
Practice the procedure of MIC test
DL:
Prepare absorptive layer by different concentrations of alginate and chitosan solution
Improve steps in the preparation process of the absorptive layer to make its structure more stable, such as pH neutralization
Swelling test
Design Wiki style
HP:
Co-product online teaching video with iGEM Team NTCU_Formosa
Pop up with an idea of the iASK symposium with iGEM Team NCKU_Tainan
Translate parent-child picture book into Kazakh and French version, thanks to iGEM Team NU_Kazakhstan and iGEM Team Aix-Marseille
Rehearsal online courses for Consistent Children Care organization collaboration
Week12: 07/19-07/25
WL:
Organize the protocol of cloning
Organize evidence supporting the severity of MRSA latency
Design the sequence of cathepsin S site
Cloning eGFP-TAT and eGFP into expression vectors- ligation into expression vectors
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP, and thrombin releasable AMP into expression vector- ligation into expression vectors
Practice the procedure of MIC test
DL:
Prepare absorptive layer by different concentrations of alginate and chitosan solution
Swelling test
Design Wiki style
HP:
Typeset our picture book
Prepare the iASK symposium with iGEM Team NCKU_Tainan
Contect St. Viator Catholic High School
Week13: 07/26-08/01
WL:
Explore the stability of antimicrobial peptides in a low pH value environment
Explore the mechanism that S. aureus resistant lysosomal enzyme
Organize evidence supporting the severity of MRSA latency
Cloning eGFP-TAT and eGFP into expression vectors- ligation into expression vectors
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP, and thrombin releasable AMP into expression vector- ligation into expression vectors
Practice the procedure of MIC test
DL:
Prepare absorptive layer by different concentrations of alginate and chitosan solution
Swelling test
Protein structure prediction
Build the network structure of the simulation model
HP:
Have an online meeting with iGEM Team IISER Kolkata
Translate iGEM Team Ioannina’s promotion video into Mandarin Chinese
Upload 1st episode of our soap opera “We Don’t Like Germs.”
August
August
Week14: 08/02-08/08
WL:
Explore the stability of antimicrobial peptides in low pH value environment
Explore the mechanism that S. aureus resistant lysosomal enzyme
Organize evidence supporting the severity of MRSA latency
Cloning eGFP-TAT into expression vectors-ligation into expression vector
Cloning eGFP into expression vectors-protein induction
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP, and thrombin releasable AMP into expression vector- ligation into expression vectors
Practice the procedure of MIC test
DL:
Analyze the result of swelling test
Combination of collagen layer and absorptive layer
Confirm the experiment information of thrombin cleavage assay
Infer the differential equation from reaction rate equation
HP:
Operate the picture book readers survey
Participate in a campaign with iGEM Team Aix-Marseille
Collaborate with iGEM Team USP-BR
Week15: 08/09-08/15
WL:
Explore the mechanism that S. aureus resistant lysosomal enzyme
Cloning eGFP-TAT into expression vectors-ligation into expression vector
Cloning eGFP into expression vectors-protein induction
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP, and thrombin releasable AMP into expression vector- ligation into expression vectors
Practice the procedure of MIC test
Stimulate the monocyte to macrophage
DL:
Combination of dressing
Use differential equation and law of conservation of mass to built the framework simulation model
Cathepsin S cleavage preference prediction
HP:
Have an online meeting with iGEM Team Tübingen
Launch a picture book promotion collaborate with Teach For Taiwan
Collaborate with Consistent Child Care
Upload online science courses to LearnMode and YouTube
Interview with Yien Home Care Center
Week16: 08/16-08/22
WL:
Find out that whether the mechanism of S. aureus resistant lysosomal enzymes impacts Cathepsin S
Cloning TAT-GFP and eGFP into expression vectors-Protein induction
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP, and thrombin releasable AMP into expression vector- ligation into expression vectors
Practice the procedure of MIC test and hemolysis
DL:
Use MATLAB to built a simulation model
Confirm the active site of Cathepsin S
Conduct the docking through AutoDock Vina
HP:
Interview with Associate Professor Chien-Chi Liu
Cohold iASK symposium with iGEM Team NCKU_Tainan
Record Podcast with Aiden Lo
Conduct a dressing market survey
Prepare for the final video
Join in the SDGs impact challenge launched by iGEM Team TAS_Taipei
Upload the online science course video
Week17: 08/23-08/29
WL:
Explore the Interaction between AMP and antibiotics
Ensure our antimicrobial agent won’t irritate the wound
Cloning TAT-GFP and eGFP into expression vectors-Protein purification
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP, and thrombin releasable AMP into expression vector- ligation into expression vectors>
Cloning CBD-linker-thrombin cleavage site for buforin II-C mimic release-GFP into expression vector- protein induction>
Practice the procedure of MIC test, hemolysis, and cytotoxicity>
DL:
Improve the dressing combination method by coating the collagen layer on the absorptive layer
Use MATLAB to built a simulation model
Confirm the substrate binding site on Cathepsin S via crystal structure
HP:
Upload the third episode of the soap opera
Interview with Dr. Chi, Chia-Yu
Have an online meeting with iGEM Team Korea_HS
Have an online meeting with iGEM Team IISER Kolkata
Have a picture book promotion collaborate with Concordia Kindergarten
Complete the online science video with iGEM Team NCTU_Formosa
September
September
Week18: 08/30-09/05
WL:
Explore the pH-activate intein
Cloning eGFP-D2A21 into expression vectors-Colony PCR and restriction enzyme digestion
Cloning CBD-linker-thrombin releasable AMP into expression vectors-Restriction enzyme digestion and protein induction
Cloning CBD-linker-thrombin releasable TAT-DPK-060 into expression vectors-protein induction
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP into expression vector- Ligate into pRSET vector, Colony PCR, protein induction
Cloning CBD-linker-thrombin cleavage site for buforin II-C mimic release-GFP into expression vector- protein induction- Time course, cell disruption
Perform of MIC and hemolysis
DL:
Ligand preparation
Confirm the specificity pocket of CTSS
Combination of three-layer dressing
Dressing & target population analysis
Data analysis via ImageJ
HP:
Prepare for the interview with the Industrial Technology Research Institute (ITRI)
Prepare for the interview with Dr. Lee, Nan-Yao
Week19: 09/06-09/12
WL:
Explore the Anti-biofilm ability of AMP
Perform cell penetration ability test
Cloning CBD-linker-thrombin releasable D2A21 into expression vectors-protein induction
Cloning CBD-linker-thrombin releasable TAT-DPK-060 into expression vectors-protein induction, Time course
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP into expression vector-protein induction, cell disruption
Perform of MIC, hemolysis, and cytotoxicity
DL:
Ligand preparation
Protein-ligand docking through AutoDock
Analyze the pros and cons of dressing combination method
HP:
Interview with experts from ITRI
Interview with Dr. Lee, Nan-Yao
Visit Trade Wind Biotech Co., Ltd
Record Podcast with iGEM Tübingen
Collaborate with Dong Rong Elementary School
Upload the fourth episode of the soap opera
Week20: 09/13-09/19
WL:
Perform cell penetration ability test
Cloning CBD-linker-thrombin releasable D2A21 into expression vectors-protein induction, Time course, cell disruption
Cloning CBD-linker-thrombin releasable TAT-DPK-060 into expression vectors-protein induction, watson blot
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP into expression vector-protein induction, cell disruption in various condition, watson blot
Perform of MIC, hemolysis, and cytotoxicity
DL:
Practice the procedure of thrombin cleavage assay
Data analysis via ImageJ to get the release curve
Use MATLAB to built a simulation model
Protein-ligand docking through AutoDock
HP:
Shoot our team presentation video
Interview with Associate Professor Jong-Yuh Cherng
Participate in iGEM TAS_Taipei SDGs Challenge Conference
Online program with St. Viator Catholic High School
Week21: 09/20-09/26
WL:
Cloning CBD-linker-thrombin releasable AMP into expression vectors-Colony PCR, protein induction
Cloning CBD-linker-thrombin releasable TAT-DPK-060 into expression vectors-cell disruption
Cloning CBD-linker, thrombin cleavage site for AMP mimic release-GFP into expression vector-cell disruption, small scale protein purification
Cloning CBD-linker, thrombin cleavage site for TAT mimic release-GFP into expression vector-Colony PCR, Restriction enzyme digestion
Perform cytotoxicity
DL:
Protein-ligand docking through AutoDock
Practice the procedure of thrombin cleavage assay
Data analysis & fitting (MATLAB)
HP:
Participate in Innovation and Entrepreneurship Competition
Design a flyer on using antibiotics accurately
Have an interview with the iGEM Guelph team
Week22: 09/27-10/03
WL:
Perform cell penetration test
Cloning CBD-linker-thrombin releasable AMP into expression vectors-Colony PCR, small scale protein purification
Cloning CBD-linker-thrombin releasable TAT-DPK-060 into expression vectors-small scale protein purification
Cloning CBD-linker, thrombin cleavage site for AMP mimic release-GFP into expression vector-time course, small scale protein purification
Cloning CBD-linker, thrombin cleavage site for TAT mimic release-GFP into expression vector-Colony PCR
Perform hemolysis and cytotoxicity
DL:
Normalize the picture of PAGE via MATLAB
Data analysis & fitting via MATLAB
Protein-ligand docking through AutoDock
HP:
Organize silver dressing information based on Dr. Lee’s feedback
Interview with Dr. Tak-Wah Wong
Contact with local dermatology clinics
Write subtitles for two videos
Edit the video interviewing foreign students in CCU
Design the slides of the presentation video
Edit the presentation video
October
October
Week23: 10/4-10/10
WL:
Cloning CBD-linker-thrombin releasable TAT-DPK-060 into expression vectors-Cell disruption, FPLC
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP into expression vector-protein induction, cell disruption, small scale protein purification
Perform MIC, hemolysis, and cytotoxicity
DL:
Integrate the data and put it on the wiki
Use MATLAB to built a simulation model
Protein-ligand docking through AutoDock
Analyze gel of SDS-PAGE via normalization between different gel images and signal integration of each band
Conduct thrombin cleavage assay
Data analysis & fitting via MATLAB
HP:
Upload the last episode of the soap opera
Film and edit the presentation video
Subtitle the video interviewing foreign students in CCU
Week24: 10/11-10/17
WL:
Perform cell penetration test
Cloning CBD-linker-thrombin releasable TAT-DPK-060 into expression vectors-FPLC
Cloning CBD-linker, thrombin cleavage site for AMP/TAT mimic release-GFP into expression vector-ligate to pET32a vector, Colony PCR, FPLC
Perform MIC and cytotoxicity
DL:
Integrate the data and put it on the wiki
Use MATLAB to built a simulation model
Protein-ligand docking through AutoDock
Analyze gel of SDS-PAGE via normalization between different gel images and signal integration of each band
