Team:EPF Lausanne/Calendar/27 September 2013

From 2013.igem.org

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''New Strategy'' <BR>
''New Strategy'' <BR>
- We found an overlapping between the primers so this could explain why the cloning doesn't work.  
- We found an overlapping between the primers so this could explain why the cloning doesn't work.  
 +
<br>
''Western Blot'' <BR>
''Western Blot'' <BR>
- As the last WB didn't work we repeated the experiment keeping the same samples but making carefully the calculation to make the gels.
- As the last WB didn't work we repeated the experiment keeping the same samples but making carefully the calculation to make the gels.
<br>- This time the migration and the transfer went well. But we still don't have a good positive control.
<br>- This time the migration and the transfer went well. But we still don't have a good positive control.
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<br><br>
<font size = "4"> Sensing-Effector </font> <BR>
<font size = "4"> Sensing-Effector </font> <BR>
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''Measuring GFP expression'' <BR>
''Measuring GFP expression'' <BR>
-Again I tried to induce GFP expression in the bacteria containing either the hya or the cad promoter.  
-Again I tried to induce GFP expression in the bacteria containing either the hya or the cad promoter.  
-
This time I washed and dilluted the cells before putting them in the different media. I also made three replicates of each medium, i.e. three wells with pH 5.5, three well with pH 7, etc.
+
This time I washed and diluted the cells before putting them in the different media. I also made three replicates of each medium, i.e. three wells with pH 5.5, three well with pH 7, etc.
The absorbance was measured in a plate reader during 18h, a measurement was taken every 30 minutes.
The absorbance was measured in a plate reader during 18h, a measurement was taken every 30 minutes.
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Latest revision as of 22:28, 4 October 2013

Taxi.Coli: Smart Drug Delivery iGEM EPFL

Header

Cell Surface Display
New Strategy
- We found an overlapping between the primers so this could explain why the cloning doesn't work.
Western Blot
- As the last WB didn't work we repeated the experiment keeping the same samples but making carefully the calculation to make the gels.
- This time the migration and the transfer went well. But we still don't have a good positive control.

Sensing-Effector

Measuring GFP expression
-Again I tried to induce GFP expression in the bacteria containing either the hya or the cad promoter. This time I washed and diluted the cells before putting them in the different media. I also made three replicates of each medium, i.e. three wells with pH 5.5, three well with pH 7, etc. The absorbance was measured in a plate reader during 18h, a measurement was taken every 30 minutes.

Human practice
Kit
- Remaking the transformation starting with NEB 5-alpha high efficiency competent cells.