Document

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 lab activity this week, University Final 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!!

Clinging a little bit longer to summer vacations.

Actualization meeting after holidays, we start to search for financial support.

Presentation to CeBiB asking for financial and academic support.
Stay selling in the University for friday parties.

Attending to the MIT Global Start-Up Workshop, for learning how to do a correct elevator pitch.

Meeting with professor Franck Quero, we chat about types of cellulose and more of its wordeful properties

Working on Franck Quero's lab, growing some G. xilinum.

Collecting our bacterial cellulose!!!

No activity in lab this week

Lauching our online Survey

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.

Participation on the radio show "I want to be scientific" ("Quiero ser científico") with high school students.
Exposition of the project to kids in the science fair held at Ñuñoa square.

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.

Discussion about the answers made in the las meeting and the mathematical model

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.

Modellating meeting, looking for the parameters from Brenda

We launched our crowdfunding video! you can watch it if you like

Our first and longer collaboration started with iGEM Pasteur

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.

Meeting with Lucho :D , Some modellating problems are solved and was proposed for the next meeting, have the most efficient promoter

We improved the genetic circuit. Maymi accepted to be part of the team.

Find out that it is almost impossible to synthesize bacterial cellulose, K.O. cellulose production :(

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.

First aproach to design our poster!!!

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.

Meeting in the Energy Ministry

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.

Workshop on the XI Science and Technology Fair in the Metropolitan park, we have kids visiting us!!!

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.

Visit to Los Maitenes and Focus Group with the neighbors

Chemical and Biotechnological Week were we explained what is a bioreactor and lauch a contest.

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

Attendance to the VI National Congress of Biotechnology held in Concepción

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.

VERY HARD WORK in wiki

Team:UChile OpenBio-CeBiB/Notebook

Notebook