Team:EPF Lausanne/Calendar/15 September 2013

From 2013.igem.org

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{{Template:EPFL2013Header}}
{{Template:EPFL2013Header}}
 +
<font size = "4"> Cell Surface Display </font> <BR>
-
Sensing <BR>
+
''New Strategy'' <BR>
 +
- Optimization of PCR using the new primers designed to amplify the different component that will be needed to clone the following constructs : INP_YFP_Streptavidin and INP_streptavidin_YFP.
 +
<br>- Beginning of gibson reactions involving the previously amplified DNA sequences.
 +
<br>- PCR to amplify Streptavidin without a stop codon needed for the INP-Streptavidin-YFP construct. Gel showed, that nothing was amplified. Repeated the assay with a different PCR programm. Gel showed expected bands.
 +
<br>
 +
''Inoculation for Western Blot'' <BR>
 +
Overnight inoculation of ISA, ISI, ISD, Streptavidin initial plasmid transformed cells and competent cells.
 +
<br><br>
-
'''Analysis of the sequencing resutls of the three promoters (sensing-module)''' <BR>
+
<font size = "4"> Sensing-Effector </font> <BR>
-
The sequencing results were rather promising except for the Hya promoter. The sample that I send did not contain it. So I purified one in order to send it again on Tuesday. This sample was from one of the assays. I send this sample because it did not turn green when I added arabinose indicating that the Arabinose inducible promoter had effectively been removed and hopefully replaced by the hya promoter.
+
-
'''Functional assays for the three promoters (sensing-module)'''<BR>
+
''Analysis of the sequencing results of the three promoters (sensing-module)'' <BR>
-
I again tried to grow the bacteria in an LB medium to which I added different amounts of either 10X MOPS or 10X HEPES buffer. But again the cultures did not turn green.
+
-The sequencing results were rather promising except for the Hya promoter. The sample that I send did not contain it. So I purified one in order to send it again on Tuesday. This sample was from one of the assays. I send this sample because it did not turn green when I added arabinose indicating that the Arabinose inducible promoter had effectively been removed and hopefully replaced by the hya promoter.
 +
''Functional assays for the three promoters (sensing-module)''<BR>
 +
-I again tried to grow the bacteria in an LB medium to which I added different amounts of either 10X MOPS or 10X HEPES buffer. But again the cultures did not turn green.
-
'''MinPrep of the hya-plasmid''' <BR>
+
''MinPrep of the hya-plasmid'' <BR>
-
Because the sequencing results showed that the plasmid I sent did not contain the hya promoter I picked another colony that I incubated and from which I then isolated the plasmid again by a MiniPrep.
+
-Because the sequencing results showed that the plasmid I sent did not contain the hya promoter I picked another colony that I incubated and from which I then isolated the plasmid again by a MiniPrep.
 +
<BR><BR>
 +
<font size = "4"> Nanoparticles</font> <BR>
 +
''Digestion assays''<BR>
-
'''Nanoparticles'''
+
''Culture of MMP2 and gelE from Sensing group''<BR>
-
<BR>- Digestion assays
+
-
<BR>- Culture of MMP2 and gelE from Sensing group
+
{{Template:EPFL2013Footer}}
{{Template:EPFL2013Footer}}

Latest revision as of 22:50, 4 October 2013

Taxi.Coli: Smart Drug Delivery iGEM EPFL

Header

Cell Surface Display

New Strategy
- Optimization of PCR using the new primers designed to amplify the different component that will be needed to clone the following constructs : INP_YFP_Streptavidin and INP_streptavidin_YFP.
- Beginning of gibson reactions involving the previously amplified DNA sequences.
- PCR to amplify Streptavidin without a stop codon needed for the INP-Streptavidin-YFP construct. Gel showed, that nothing was amplified. Repeated the assay with a different PCR programm. Gel showed expected bands.
Inoculation for Western Blot
Overnight inoculation of ISA, ISI, ISD, Streptavidin initial plasmid transformed cells and competent cells.

Sensing-Effector

Analysis of the sequencing results of the three promoters (sensing-module)
-The sequencing results were rather promising except for the Hya promoter. The sample that I send did not contain it. So I purified one in order to send it again on Tuesday. This sample was from one of the assays. I send this sample because it did not turn green when I added arabinose indicating that the Arabinose inducible promoter had effectively been removed and hopefully replaced by the hya promoter.

Functional assays for the three promoters (sensing-module)
-I again tried to grow the bacteria in an LB medium to which I added different amounts of either 10X MOPS or 10X HEPES buffer. But again the cultures did not turn green.

MinPrep of the hya-plasmid
-Because the sequencing results showed that the plasmid I sent did not contain the hya promoter I picked another colony that I incubated and from which I then isolated the plasmid again by a MiniPrep.

Nanoparticles

Digestion assays

Culture of MMP2 and gelE from Sensing group

Wiki Links

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