Difference between revisions of "Team:UChile OpenBio-CeBiB/Notebook"
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<button class="accordion"><p id="panelTitle">Week 18 [02/13/17]</p></button> | <button class="accordion"><p id="panelTitle">Week 18 [02/13/17]</p></button> | ||
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<button class="accordion"><p id="panelTitle">Week 19 [02/20/17]</p></button> | <button class="accordion"><p id="panelTitle">Week 19 [02/20/17]</p></button> | ||
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| − | <p> | + | <p>Holly Holidays!!</p> |
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<button class="accordion"><p id="panelTitle">Week 20 [02/27/17]</p></button> | <button class="accordion"><p id="panelTitle">Week 20 [02/27/17]</p></button> | ||
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Revision as of 15:30, 1 November 2017
Notebook
Presentation of 5 projects in the Engineering and Science Festival.
Workshops are made daily for high school students and families, aprox 300 peoples were in our Workshops!!
Presentation of the projects to the academic body of the Chemical and Biotechnological Enginnering Department.
Project selection, Greenhardtii was elected above the other 4 projects
First work meeting, selecting an appropiate organism capable of doing photosynthesis.
First presentation of the project to public.
Discussion of a possible production of a biomaterial, team is divided on 3 commisions to search for biomaterials.
The team find out fatty acids, cellulose and natural inks. The commisions previously formed investigate possible production of each biomaterial.
Presentation of the methabolics routes in the production of the biomaterials, cellulose is elected as the most potentialy produced biomaterial.
No activity in lab this week, University Exams.
No activity in lab this week, University Exams.
Meeting to form new commisions for working during holidays, Cellulose production and CO2 capture optimization.
No activity in lab this week, University Exams.
Holidays!!!
Cellulose comission investigate bacterial cellulose properties.
Cellulose comission arrange a pizza meeting with José Duguet, so we could develop a regulation system for production.
Sofi has joined the party
Cellulose comission finds the inducible promoters and investigate them.
Presentation of the research done during January.
Holly Holidays!!
Holly Holidays!!
Holly Holidays!!
Holly Holidays!!
No activity in lab this week
No activity in lab this week
No activity in lab this week
No activity in lab this week
No activity in lab this week
No activity in lab this week
No activity in lab this week
No activity in lab this week
No activity in lab this week
No activity in lab this week
No activity in lab this week
No activity in lab this week
Meeting with the team in order to determinate the list of protocols that we were going to get for the later lab work.
No activity in lab this week
It is possible to elaborate a first document with the experimental strategy and most of the protocols. It was presented to professor Álvaro, who suggested us to correct some details and contact Dr. Tomás egaña, Engineer in Molecular Biotechnology, expert working with the microalgae Chlamydomonas reinhardtii.
Reunion with the team to find the answers of the doubts about the document with the elaborated protocols.
Meeting with Dr. Tomás Egaña, who kindly helped us solve our doubts about our experimental strategy and some protocols related to Chlamydomonas reinhardtii. Finally, he talked us about Myra Chávez, a postdoc and an expert working with Chlamydomonas reinhardtii, who could share with us transformation plasmids, and other materials.
We achieved to make contact with Myra Chávez, who offered us help to solve some problems and she kindly gave us pBC1-CrGFP. From now on, Myra became a big help and support for our team and we nicknamed her as Maymi.
No activity in lab this week
Meeting to work in the sequences that must be sent to print, define, and analyze. We also searched the tools of optimization of codons for Chlamydomonas reinhardtii to make the first order to IDT.
We improved the genetic circuit. Maymi accepted to be part of the team.
Usage of the IDT tool to make the codon’s optimization.
Collaboration from Germany: LMU München (Ludwig Maximilians Universität München), they helped us check our experimental strategy and sequences design.
Meeting with Jose Duguet, who give us information about how to make the iGEM parts and helped us with some questions. We then worked in the parts design.
Meeting with professor Álvaro, design of the primers, and consultations.
Collaboration from England, Dr. Allison Smith from the Cambridge University. She shared us the promoter MetE sequence, 5’UTR and its terminator. She also gave us some advices for the elaboration of our construct.
Finished the primers design.
The sequences were finally ready to be sent to IDT.
The shipment of the sequences was made to IDT.
Meeting with Maymi and professor Álvaro to specify the experimental strategy, materials and required methods.
Amplification of pBC1-CrGFP in E. coli DH10B chemically competent.
Repetition of the last lab. Because there was no growth on the plate, the amplification was repeated. However, now the transformation was made in the E. coli top 10 chemically competent strains with more care and caution. After a thermic shock and incubation TB medium for an hour and a half, the cells were transfered to several plates with LB and ampicillin, and they were left incubating at 37°C.
The plates were checked and there actually was presence of transformant colonies with resistance to ampicillin, two of them were inoculated in Falcon tubes with LB and ampicillin medium at 37°C.
Miniprep of the transformants colonies in Falcon. Isolation of the plasmid pBC1-CrGFP.
Test transformation of the Chlamydomonas reinhardtii by method of glass beads with pBC1-CrGFP. We also learnt to cell count on a Neubauer camera. After the transformation the cells were left incubating in Falcon tubes with TAPS medium in dark during the night at room temperature with agitation.
Continuing with the last lab. The cells were incubated in TAPS, then they were centrifuged and sowed in plates with TAPS medium with agar and paromomycin. They were left incubating at ambient temperature.
No activity in lab this week
Due to that we were informed that some of the sequences that were sent could not be printed, a second order was made to IDT with the correct sequences.
Arrival of the first order from IDT.
Resuspension of the lyophilized gBlocks.
It was informed to us that one of the sequences from the second order could not be synthesized again.
Resuspension of the universal primers and PCR of the gBlock of the first order from IDT.
Preparation of gel extraction of amplified sequences cut DNA bands.
Extraction and purification of DNA from the agarose gel bands.
Digestion of the standard sequences and the igem vector pSB1C3. Ligation with the standard sequences.
Transformation by electroporation of the vectors and sequences in BL21DE3 E. coli strain. Seeding in selective mediums, they were left incubating at 37°C until 2 p.m. of the next day.
Isolation of the colonies and growth in LB liquid medium with antibiotic. Left at 4°C
Miniprep of the colonies and generation of a glycerol inventory.
First test of miniprep digestion. The samples were saved at -20°C.
Preparation of visualization gel of the miniprep digestion. Bands on the gel were not observed.
The visualization gel was repeated and again bands on the gel were not observed. The part was sent.
We decided to repeat the test of miniprep digestion and electrophoresis was carried out. Finally, we were able to observe the bands, which validated the construct with the part that was sent.
No activity in lab this week
ono
ono
