Team:TU-Eindhoven/Notebook/Week8

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• Introduction
  • Home page
  • Background
    information
  • About us
  • Contact us
  • Our photos
• Project
  • Project description
  • Lab approach
  • Device information
  • Initial model
  • Yeast model
  • Lab results
  • Our BioBricks^TM
  • Our
    accomplishments
• Considerations
  • Public outreach
  • Safety questions
  • Other thoughts
• Future applications
  • Human interest
  • Further
    developments
• Notebook
  • Before week 1
  • Week 1
  • Week 2
  • Week 3
  • Week 4
  • Week 5
  • Week 6
  • Week 7
  • Week 8
  • Week 9
  • Week 10
  • Week 11
  • Week 12
  • After week 12
  • Lab protocols
• Partners
  • ICMS TU/e
  • Team collaborations
  • Attributions and Contributions
  • Our sponsors

The mock-up for our presentation is finished and looks marvelous, thanks to the ICMS Animation Studio! Besides this great news, there was also sad news. The iGEM tem from Delft quit organizing the Discovery Festival. The remaining four Dutch iGEM teams (from Groningen, Wageningen, Amsterdam and Eindhoven) are now working extra hard to accomplish our ideas.

The PCR-step and gel extraction for the BioBricks had to be redone because of mixed up microcentrifuge tubes. Furthermore, the restriction and ligation for the BioBricks are made. We made chlorampenicol plates and we transformed the Nova Blue E. coli with the ligation mix to obtain colonies carrying the BioBricks. But since we didn’t place the plates containing E. coli upside down, they dried out and the transformation failed. Therefore the transformation is repeated. Unfortunately, the BioBrick assembly of pSBIC3+GECO failed. We couldn’t find any colonies on the plates and in the bottles. To check the PCR product we used, we tested it on a gel. The gel showed that we got the right inserts. The problem seemed to be the ligation and restriction. A new PCR product of the GECOs is made as preparation for the assembly.

Luckily, the results of the yeast transformations are positive. The single-plasmid transformations worked out well, the co-transformations however were not successful, except for the Mid1+Cch1-EGFP. The medium for another co-transformation was made, but one of the required ingredients for the medium didn’t solve. Therefore we had to make this type of medium again by another method. We picked colonies from the yeast plates and made 5ml O/N cultures and a 5ml InvSC1 with R-GECO preculture. Unfortunately, we had to deal with some small disturbances this week. The fire alarm went off, so we had to leave the building for fifteen minutes, we spilled some yeast in the incubator, so we had to clean up for another fifteen minutes, and so on…

We tested the device, which seemed to work technically. Little bubbles occurred in the medium after activating the device. But still no fluorescent light is visible.