Team:UC Chile/Cyano/Notepad/week20

From 2012.igem.org

Monday: Today is suppose to be a holiday, I'm not really sure why, but the team is still working. The building seems abandoned and we are the only ones in here muajuajuajuajua (evil laughter).

About our work in progress: we have our second arduino project. It is a photoresistor able to measure the intensity of light inputs. The arduino program is linked to old Juano's c# aplication so we can see a graph of the data in real time. We have only tried it with LEDs so far and the system proved to be sensible enough. The next step is to measure bacterial biolumniscence. We hope to do so tomorrow. We transformed e. coli with lux brick (at race speed) in order to have a really nice shining flask and check if our arduino system can sense bacterial produced light properly.

Meanwhile in the wetlab... We built the following parts trough a ligation protocol. Then we transformed and plated e.coli.

1. RS1 + KanR + psb1C3 . Plate resistance: chloramphenicol and kanamycin.
2. RS1 + KanR. Plate resistance: chloramphenicol.
3. RS1+ KanR. Plate resistance: chloramphenicol and kanamycin.
4. B0014 + RS2 + psb1C3. Plate resistance: chloramphenicol.
5. B0014 + RS2. Plate resistance: kanamycin.

Also, we did an electrophoresis run of LuxCDEG PCR product. Nothing showed up u.u. But we could purify more psB1C3 (it does not make up for lux though)

Finally, we prepared some parts to send to our international advisor (yeah, the Argentinian) so he can try to assemble them by the Gibson technique.

SIMON LEFT US TODAY :_( He'll be spending the next three (yes, THREE!) weeks in San Francisco. He couldn't help but trade us for sunny californian beaches...

Tuesday: We had a videoconference with Copenhagen and Valencia teams. As we are working in very similiar projects we decided to establish a collaborative alliance. Eventually, we will share our constructs to compare their performance in the different systems. Go teams go!!!

E. colis with lux brick were grown in LB media. L-Arabinose 2 mM was added. The sensitivity test for the arduino circuit will be done tomorrow.

Later on we checked yesterday's plates and all of them had colonies (yay!!), although the ones with newer ligation buffer had better yield (plates 1 and 5). The previous low transformation efficiencies might have been because of this. We culture colonies from plates 1 and 5 in bacterial tubes for further extraction.

Another gel run for LuxCDEG PCR product was done. Again, no sign of strand. Run for LuxCDEG with X+P also showed nothing. We think we must have done something incorrectly. We'll try the following ligation: (Lux CDEG1 E+S digestion) + (Lux CDEG2 X+P digestion) + (psB1K3 E+P).

As a day with no transformations is a dull one we transformed: Lux CDEG in psB1C3, Lux CDEG in psB1A2 and Lux CDEG in psB1A2 (from plasmid).

Checking final details of synechocystis transformation protocol. We hope to do it on Thursday!

Parts for our Argentinian advisor are on their way to Cambridge :) hope they make it through customs!

Wednesday: