IGEM:UNAM-Genomics Mexico/2009/Notebook/Collaborations/2010/10/18: Difference between revisions
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== | ==Asking for advices with co-transformations and bioluminiscence measurements== | ||
Hola Maria, | |||
We are also having problems with the spectro-fluorimeter that is available | |||
in our center of research. We can't detect the light output of the | |||
LuxABCDE genes from vibrio fischeri that Cambridge sent us; even the | |||
bioluminescence it is also visible to the naked eye. We tried the | |||
measurements after spin-down several milliliters of the culture (10, 5 and | |||
3 ml) and re-suspending, in order to get cells more concentrated, but it | |||
didn´t work. Please, share us your experience with this issue if you and | |||
Will get better results. | |||
Maybe, we will try to do the measurements with a CCD-camera that is | |||
available at the physics center, but it is a little bit complicated to get | |||
in touch with the responsible person. | |||
The co-transformation of LuxY with LuxABCDE genes has failed, though the | |||
plasmids have different replication origins; we will try to get it again | |||
along this week. | |||
I wanted to ask you, How long the bioluminescence is conserved in your | |||
cultures and Petri-dishes? Do you have any limitation by the temperature | |||
growth conditions? | |||
On the other hand, hopefully we could test LovTAP. It seems that a read | |||
out system (trpL promoter + RFP has been ligated correctly). It will just | |||
depend on the co-transformation success that we have. | |||
Do you have any advice about co-transformations; Martha told me that she | |||
had tried some, working with LovTAP. How was the experience? | |||
About the shipment, we totally respect and understand Dr. Chris position. | |||
BTW the shipment has not arrived yet. | |||
Well, Good luck during these weeks. | |||
Regards, | |||
Claudia | |||
Revision as of 22:07, 17 October 2010
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Asking for advices with co-transformations and bioluminiscence measurementsHola Maria, We are also having problems with the spectro-fluorimeter that is available in our center of research. We can't detect the light output of the LuxABCDE genes from vibrio fischeri that Cambridge sent us; even the bioluminescence it is also visible to the naked eye. We tried the measurements after spin-down several milliliters of the culture (10, 5 and 3 ml) and re-suspending, in order to get cells more concentrated, but it didn´t work. Please, share us your experience with this issue if you and Will get better results. Maybe, we will try to do the measurements with a CCD-camera that is available at the physics center, but it is a little bit complicated to get in touch with the responsible person. The co-transformation of LuxY with LuxABCDE genes has failed, though the plasmids have different replication origins; we will try to get it again along this week. I wanted to ask you, How long the bioluminescence is conserved in your cultures and Petri-dishes? Do you have any limitation by the temperature growth conditions? On the other hand, hopefully we could test LovTAP. It seems that a read out system (trpL promoter + RFP has been ligated correctly). It will just depend on the co-transformation success that we have. Do you have any advice about co-transformations; Martha told me that she had tried some, working with LovTAP. How was the experience? About the shipment, we totally respect and understand Dr. Chris position. BTW the shipment has not arrived yet. Well, Good luck during these weeks. Regards, Claudia
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