Team:UC Chile/Cyano/Notepad/week19

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Cyanolux & SpiderColi - Pontificia Universidad Católica de Chile, iGEM 2012



notebook



Monday: Well, today was our first group meeting after the final exams so yes, most of us seemed to be taken from a "The Walking Dead" set, but still none of us fell asleep (great achievement team!). Emi couldn't make it physically because she's still recovering from mononucleosis but she was present thanks to google's Hang Out.

During the meeting...

Basically we defined guidelines to follow and tasks to complete before the wiki freeze. The next two weeks were planned according to who is going to leave for holidays (Simon decided to spend all is money in Las Vegas, Carla is going to drink as much beer as she can at her homeland, Max will be off searching for white walkers way beyond the wall) and the rest of the slave... I mean team members that will be staying in Santiago therefore in the lab.

So, Simon will transfer all of his wet lab knowledge to Isaac and Tama, meaning they are going to be in charge of the ligations and restrictions for the next three weeks. Max will be reading dusty books and ancient rotten scrolls (or the newest issues of Science, Nature and others might also help) in order to find a new protein secretion system for spider silk. He'll be replaced by Ulises starting from July 23. All arduino related work will be under Carla's command, once she leaves Arduino will be in stand by (just for one week only).

Bryon needs to get results in his other lab ASAP so we won't see him around much. Seba will be studying for his big B.S. degree exam so he is in the same status as Bryon (not around much). Claudia is finishing her M.S. thesis so... I really don't know. Emi, emi emi... please get better soon!!! She'll be doing as much work as she can while in bed, most will be wiki material.

And that leaves us Pollak... :) lab work 24x7. I just hope you get to eat something.

Crazyengineer42 will be in charge of wiki as usual ;)


THE ARDUINO QUEST

Short story on how Pollak and Carla crossed half Santiago to get the Arduino kit.

16.58: Carla receives mail from Olimex manager on how to get the payment and shipping ready.
17.01: Carla forwards e-mail to Pollak.
Some time between 17.01 and 17.40: Pollak gets University's ID number and cash and heads to Olimex office. Closing time is 18.00. Carla finds out Pollak is leaving after her mid afternoon coffee. Carla joins Pollak.
Some time between 17.40 and 17.59: Carla and Pollak go by Santiago Underground to Olimex. Coach is partly crowded.
Once destination is reached, Carla and Pollak run to Olimex office. Carla and Pollak are outside the door at 17.59. Some time between 17.59 and 18.30: Carla and Pollak receive the Arduino Uno and several resistors, mini protoboards and other necessary stuff. However, the light sensor is not in stock and will be available two weeks from now.
Some time between 18.30 and 19.00: The journey back home. Underground is collapsed. Carla and Pollak get in somehow. Carla is crushed by all the people trying to protect the Arduino. Arduino is safe. Arrival time at lab around 19.00.

Tuesday: Day started off with the introduction of our newest advisor Juan Eugenio Venegas (from now on Juano). He'll be supporting us with all electronic and Arduino related issues. Juano gave a flash training to Carla on the Arduino basics like how to turn on/off a led and coding. Finally, Juano tried the c# application he created to graph data input in real time. It worked just fine, but as the sensor is not here yet a potentiometer was used instead.

A vector PCR was done to get suffient amount of pSB1C3, pSB4K5 and pSB1A2. This plasmids will be used in the ligations of lux AB and lux CDGE. Also, amplicon "psaAB-RFP" was purified with a yield of 32 ng/uL.

Wednesday:

Gels were run for the vector PCR plasmids. psB1A2 (from C) and psB1C3(from pure) turned out to be positive with a yield of 38 ng/μL and 56 ng/uL repectively. Later on, plates were made to culture the plasmids.

Carla went off to buy photoresistors to use them as replacement of the missing light sensor. We hope the new arduino system will be able to sense the light at bacterial scale.

At the end of the day, Carla and Bernardo had a meeting with Rolando (the arquitect) to go over the design of the wiki and biolamp. Again, there was no agreement on the logo. Rolando is in charge of the wiki's aesthetics for now.

Thursday: this is the day crazyengineer42 (meaning me) took the day off and wandered all day long through the streets of Santiago trying to find his/herself.

Meanwhile in the lab:

Did a PCR colony protocol to check if psbAB-sfGFP was inserted in pPMQAK1. Also, did a vector PCR for psB4K5 and psB1C3.

Fail: wrong plasmids added to colony PCR. This will probably lead to dimer formation.

Friday: We have our first Arduino project running :D It's a temperature sensor that sends the message "Pollak screw u :)" to the program console if the temperature is between 20 - 30 °C. That's the twisted sort of way we demostrate our gratitude feelings towards our instructor. Also, every time the temperature drops, the message "Brace yourselves, winter is coming" is sent. (The arduino can turn off and on a LED as well but that's not a big deal!).

Wet lab: colony PCR indeed failed and the last vector PCR didn't turn out ok. Both experiments were repeated.

For colony PCR, templates at 35 ng and 7.2 ng were used for psB4K5 and psB1C3. Finally we obtained positive results!

Vector PCR failed for the third time.

It's friday friday gotta get down on friday... (I made you remember this, again!)