Team:EPF Lausanne/Calendar/28 August 2013

From 2013.igem.org

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<font size = "4"> Cell Surface Display </font> <BR>
<font size = "4"> Cell Surface Display </font> <BR>
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''Functionnal/Characterisation Assays'' <br>
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''Functional/Characterisation Assays'' <br>
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- 1ml of inoculated samples (ISA, ISD, ISI and CC) were pelleted down, resupended in PBS and incubated 1hr with anti-streptavidin FITC conjugated antibody.
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- 1ml of inoculated samples (ISA, ISD, ISI and CC) were pelleted down, resuspended in PBS and incubated 1hr with anti-streptavidin FITC conjugated antibody.
<br>- The cells were then fix or kept in liquid (see protocols) medium and visualized under the microscope.
<br>- The cells were then fix or kept in liquid (see protocols) medium and visualized under the microscope.
<br>- No fluorescence was observed.  
<br>- No fluorescence was observed.  
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''Checking GFP expression induced by arabinose'' <BR>
''Checking GFP expression induced by arabinose'' <BR>
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-We wanted to induce simple GFP expression in bacteria transformed with the AraC-GFP plasmid to check if the bacteria really did contain the plasmid and also if it worked the way it is supposed to. In order to do so, we innoculated two cultures into a chloramphenicol-Arabinose containing medium and let them grow over night.
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-We wanted to induce simple GFP expression in bacteria transformed with the AraC-GFP plasmid to check if the bacteria really did contain the plasmid and also if it worked the way it is supposed to. In order to do so, we inoculated two cultures into a chloramphenicol-Arabinose containing medium and let them grow over night.
''Resuspending K-12, MG1655 e.coli'' <BR>
''Resuspending K-12, MG1655 e.coli'' <BR>

Revision as of 22:03, 4 October 2013

Taxi.Coli: Smart Drug Delivery iGEM EPFL

Header

Cell Surface Display

Functional/Characterisation Assays
- 1ml of inoculated samples (ISA, ISD, ISI and CC) were pelleted down, resuspended in PBS and incubated 1hr with anti-streptavidin FITC conjugated antibody.
- The cells were then fix or kept in liquid (see protocols) medium and visualized under the microscope.
- No fluorescence was observed.

Sensing-Effector

Checking GFP expression induced by arabinose
-We wanted to induce simple GFP expression in bacteria transformed with the AraC-GFP plasmid to check if the bacteria really did contain the plasmid and also if it worked the way it is supposed to. In order to do so, we inoculated two cultures into a chloramphenicol-Arabinose containing medium and let them grow over night.

Resuspending K-12, MG1655 e.coli
-Today we also recieved the E.Coli strain which contained two of the three promoter that we wanted to use for our sensing module. We resuspended this strain according to the manufacturer's instructions and let them grow over night.

Nanoparticles

Digestion of the Nanoparticles - First try for the digestion with matrix metalloprotease MMP2: 2 different media were used, Tris/HCl with NaCl and CaCl2 and cell media. We did two measurements of the size of the nanoparticles: the nanoparticles became a little bit smaller in the presence of MMP2.