Team:EPF Lausanne/Calendar/27 October 2013

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<font size = "4"> Cell Surface Display</font>
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<BR>''CLoning INP_EYFP_Streptavidin (IYS) and INP_Streptavidin_EYFP (ISY)'' <BR>
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The transformation from the day before worked. There were no colonies is the insert and the competent cells control as expected.
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There were colonies in the backbones control and in the gibson transformation plates.
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<br>
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1) Colony PCR verification
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<br>- 0.8% Gel electrophoresis to verify the colony PCR of the day before.
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<br>- we obtained amplifications for colonies 1,3,5,6,7 of IYS and 1,4,6 of ISY.
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<br><br>
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2) Microscopy
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<br>- samples from IYS colonies 1,3,5,6 and ISY colonies 1,3,4,6 were prepared acording to our protocols.
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<br>- phycoerythrin conjugated anti-YFP antibody , FITC conjugated anti-Streptavidin antibody and Fitc conjugated biotin were used.
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<br>- unfortunatly the camera of the inverted microscope was not working so we could not save the images.
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<br>- bacteria samples were rescued by adding 2ml LB_Chl_IPTG them and incubated overnight for the experiment to be repeated the next day.
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<br><br>
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Latest revision as of 23:43, 28 October 2013

Taxi.Coli: Smart Drug Delivery iGEM EPFL

Header

Cell Surface Display
CLoning INP_EYFP_Streptavidin (IYS) and INP_Streptavidin_EYFP (ISY)

The transformation from the day before worked. There were no colonies is the insert and the competent cells control as expected. There were colonies in the backbones control and in the gibson transformation plates.
1) Colony PCR verification
- 0.8% Gel electrophoresis to verify the colony PCR of the day before.
- we obtained amplifications for colonies 1,3,5,6,7 of IYS and 1,4,6 of ISY.



2) Microscopy
- samples from IYS colonies 1,3,5,6 and ISY colonies 1,3,4,6 were prepared acording to our protocols.
- phycoerythrin conjugated anti-YFP antibody , FITC conjugated anti-Streptavidin antibody and Fitc conjugated biotin were used.
- unfortunatly the camera of the inverted microscope was not working so we could not save the images.
- bacteria samples were rescued by adding 2ml LB_Chl_IPTG them and incubated overnight for the experiment to be repeated the next day.

Wiki Links

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