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Team:TU-Eindhoven/Notebook/Week8
From 2012.igem.org
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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 <span class= "red">right inserts</span>. The problem seemed to be the ligation and restriction. A new PCR product of the GECOs is made as preparation for the assembly. | 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 <span class= "red">right inserts</span>. 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 <span class= "red">yeast transformations are positive</span>. 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 | + | Luckily, the results of the <span class= "red">yeast transformations are positive</span>. 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 5 ml 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... |
Revision as of 01:15, 27 September 2012
Update on our general tasks
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.
Lab results
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 5 ml 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...
Our device
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.
Bachelor thesis
Since the government of the Netherlands suddenly decided that all Dutch students need to finish their Bachelor in four years (otherwise you have to pay 3000 euros), one of our team members is slightly stressed out for finishing her thesis on the iGEM model. Luckily, her little brother was there for some support.
