Team:UNIFI/Journal

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Journal

May: We started to build the main idea of our project.
15/05 - Arrival of the iGEM kit. Full of wonderful parts with an incredible varieties of functions, giving us a great push towards our goals!
17/05 – 06/06 development of the bioinformatic part of the project. During this period we mostly constructed our plasmid so that we could order the missing parts and start with the laboratory work.
June: In this month we dedicated mainly to laboratory work to make practice in bacterical transformation in order to improve our results.
13/06 first bacterical transformation of pSB1C3 in E.coli; sadly first attempt failed.
20/06 second bacterical transformation of pSB1C3 in E.coli, IT WORKED!
July: We participated to the european meetup in Delft to expose our project and to meet all the other european teams! It has been an amazing experience and after it we started a collaboration with the CeBiTec-Bielefeld team; we decided in agreement to Skype every week so that we could learn from a much expert team as their's.
06-07-08/07 European meeting in Delft, three days weren’t enough for such a beautiful experience!
11/07 arrival of the Gibson Assembly kit containing freshly synthesized DNA fragments.
17/07 first Skype with the Bielefeld Team, they gave us a ton of tips about iGEM competition since many of its aspects were hidden to us. We actually discovered how deep is the rabbit hole!
24/07 Skype #2 with Bielefeld: they deeply explained us the function of the wiki and the complexity of building it, looks like we have a lot to think about! Also they gave us very useful advices on Human Practices, explaining us the goals and objectives of such a difficult task.
26/07 First time we tried the elettroporation as a technique to make the bacterial transformation
31/07 Skype #3 : discussion about the wiki, Maximilian suggested to avoid javascrit considering that it’s harder to handle and we haven’t any informatics in our team!
August: was all about the development of the wiki and the human practices. We received two parts from Bielefeld team's project to work upon, so that we could do something in exchange for their mentoring in the frame of the ongoing collaboration. This month our project in lab was mostly frozen until the fourth week.
07/08 Skype #4: Chris shared with us his protocol for the growth of the Escerichia coli engineered with their two plasmids; as soon as we receive them, we will be able to investigate the differences in the expression of the proteins encoded in their sequence. They also gave us some tips for our travel to Boston for the Jamboree!
21/08 Skype #5: discussion about how to develop conferences for our Human Practices, calling different professors and experts to have their opinions on our project. Also, we asked many infos on medal criterias and how to reach them.
28/08 Skype #6: Our fluorometer is pretty good and can help a lot our german friends! We can take a lot of measurements during the exponential phase in an automatic way, improving a lot their data. They gave us advices on how we could resolve our problems with PCR, since it's not working for 5 of our 9 DNA fragments!!
September: has been full of work: we reached the first good results in the lab for the Bielefeld project, and for our too. Moreover we organized two conference calls with two professors which have been very stimulating and interesting.
05/09 Skype #7: tips for the Conference call with professor Dirk Stermerding, and about how to find others experts in our country to help us with various aspect of our project, like how to handle the public engagement and the what should be considered before writing down a survey.
18/09 Skype #8: Team Bielefeld gave us various advices and tips to improve our PCR results, electrophoresis’s run and other pratical stuff. The best part is that they shared with us the protocol for aquacloning, a pretty unexpected techniques that we welcomed with arms wide open.
20/09 Skype call with Prof. Dirk Stemerding, senior researcher at the Rathenau Instituut (NL)
25/09 Skype #9: We must sadly announce that our third vector must be cut off from the project because we have tried all the possibilities, but seems like ligase won't work in any way this time. Bielefeld team shared with us a protocol to make competent cells, during the summer we had some problems with our protocol CaCl2-based due to high external temperatures (the average of max temperature was over 30°C for the whole sumer!)
27/09 First positive results for Bielefeld project. Conference call with David Kong about science art! David Sun Kong, Ph.D., is a Synthetic Biologist, community organizer, based in Lexington, MA. He is the Director of the MIT Media Lab's new Community Biotechnology Initiative.
October: This month has probably been the most intense, full with the many different activities in every aspect of the competition and we couldn't be more happy with all the progresses we've made so far.
2/10 First Meeting with an high school, located in Prato. We were a bit nervous but everything went fine. Skype #10: Brainstorming about Professor David Kong’s tips and how to apply them. The city’s blind people association could be interested to a collaboration with us, our software could give them the chance to understand lab work and sense the difference between different datas.
5/10 Acquacloning is a success! It's a versatile and simple enzyme-free cloning approach. Acquacloning solved our plasmid closure problems...After a month ! It's magic or maybe all genetic engineering is magical.
9/10 Skype #11: discussion about our project, our wiki and collaborations
12/10 Creation of vectors (e.g. pJet) and transformation into bacteria. The transformation method is electroporation.
14/10 Our team went to high school Castelnuovo in Florence to present the iGem project.
18/10 Skype #12: discussion about our medals and parts, both were complicated to deeply understand, so they checked our problems and asked their advisors too! Moreover, considering we are in the track "Art & Design", we have to pay attention to different criterias than the standard ones.
18/10 Transformation by electroporation of the our plasmid and positive control. Unfortunately, only positive control growed but not our clone. In the evening we extract our plasmid from the colonies transformed with pJet. We have a large number of copies of the plasmid in the cell → no need for pcr !
19/10 Plasmid extraction – kit Macherey Nagel .. no results :( But Pjet is a "suicidal plasmid", and since the colonies are growing, our fragments entered!
20/10 Castelnuovo High School students came to our labs to see the progress of our team. Everyone liked this day at the university as small scientists! These days our team also visited middle schools in Prato. The group of students was very heterogeneous but it was a great success!
23/10 Experiment with Tecan: The multi plate reader is used to evaluate the total fluorescence present in the sample. This instrument can read fluorescence directly from multi-well plates, a highly advantageous solution for simultaneous analyzes on different samples (genetic circuits). We finally got Fluorescence!
23/10 – 24/10 Our team went to high school itis Da Vinci to present the iGem project. Many young people attended the meeting and they were very excited about it!
30/10 Skype #13: A long lasting talk with Chris regarding how to complete the wiki with only a couple days remaining, an almost impossible task! This Skype would be the second last before Boston and the waiting is already killing us!

Team Unifi

unifi.igem@gmail.com