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Team:UC Chile/SpiderColi/Notepad
From 2012.igem.org
Week 1: 5-9 march
Tu:The whole team met for the first time after the vacations of february. We discussed our scheduling and divided ourselves in two groups: Cyano and Bactomythril. The leaders of each group were elected: Carla (Cyano) and Claudia (Bactomithril).
Cyano: Wetlab: Simon, Seba / Drylab: Carla, Tamara, Isaac
Biomithril: Wetlab: Ulises, Bryon / Drylab: Claudia, Emilia, Max
Carla and Max were chosen to maintain the wiki updated.
Bernardo and Rolando are going to collaborate with both groups.
Also, we scheduled meetings of the whole team every Friday or Tuesday (depending on the availability of the majority of the team), and meetings with the whole team and our advisor, Professor Rodrigo Gutierrez, every other Thursday. Likewise, each group should have their own meetings.
Finally, we decided to work based on delivery dates. Each member should compromise to complete a task in specific time intervals.
Week 2: 12-16 march
Week 3: 19-23 march
Week 4: 26-30 march
April
Week 1: 2-6 april
Week 2: 9-13 april
We: We have lunch-meeting where we decide to change our responsabilities.
Wetlab: Brion y Max
Modelation: Emilia y Ulises
Wiki: Claudia
Our task for next week are:
Wetlab: Work with the new material that arrived, primer and plasmid.
Modelation: Read paper Hayashi 1999, Hypotheses that correlate the sequence, structure, and mechanical properties of spider silk proteins. Learn how to use the software.
Week 3: 16-20 april
We: We have our regular lunch-meeting. Gibson Assembly of Working Plasmid Construct and transformation.
Th: Meeting with Rodrigo Gutierrez We have a colony after the transformation!!! :D
a) Removing the parts that deal to make the construct (Working Plasmid) and transforming them into Colis.
b) Miniprep of all constructs that transform and obtain DNA
c) Constructs were amplified in the thermocycler. We did PCR, but wasn’t in good condition, running buffer was weird was going very slow so we decided to leave it for next week.
Week 4: 23-27 april
Tu: Meeting with Rodrigo
We: We have a stand in Feria Ingenia
Th: We participate in a conversation table in Feria Ingenia. [http://twitcam.livestream.com/9pslw Livestream]
a) PCR amplification of the constructs. Of the 7 constructs to amplify, we can only see 5 in gel electrophoresis. Is due in large part because one of them is 42kb (HIV cleavage site) and a 2 kb (vector backbone; psB1K3) and the gel was not as resolute (1%) to observe clearly.
b) The gel was run with the 2 constructs mentioned. This time, add more of them (4UL template in PCR) to ensure that they were present in our tubes.
c) We observed 2 bands for both constructs.
d) We made the decision to leave the Gibson for next week.
May
Week 1: 1-4 may
a) It was made at 7 Gibson parts, to obtain finally the Working Plasmid (see section constructs).
b) Transform E. coli and then in culture plates (LB + KanR), we expect to grow overnight.
c) There are red plates; we don’t expect that, we think is a contamination. So again we held the Gibson plasmid construct working (on two different plates). In one observed 2 colonies grow in the other about 10 (:D!).
d) In the wake of the red colonies, we believe that Gibson could not have come out quite right. To ensure that our construct was made in good way, we construct colony PCR assembly (with / without taking into account the vector backbone). When you run the gel, we realized that our construct actually corresponded to the expected product (hence the Gibson worked OK, :D).
e) As our work Gibson, Gibson performed another to generate our second and third construct (HIV protease sfGFP reporter and producer, respectively).
f) Competent bacteria were transformed with the products of previous Gibson.
Week 2: 7-11 may
Week 3: 14-18 may
Week 4: 21-25 may
Week 5: 28 may-1 june
June
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