Team:EPF Lausanne/Calendar/8 September 2013

From 2013.igem.org

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<font size = "4"> Cell Surface display </font> <BR>
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<font size = "4"> Sensing </font> <BR>
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''Characterization/Functional Assays '' <BR>
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Inoculation of all our constructs and controls (competent cells, INP-Strepta-igem, INP-Strepta-dead, INP-Strepta-alive, BBa_K523013, YFP control, GFP control) needed for the confocal microscopy.
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<font size = "4"> Sensing-Effector </font> <BR>
''PCR of MMP9 insert and GelE insert'' <BR>
''PCR of MMP9 insert and GelE insert'' <BR>
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''DpnI digest of MMP2 insert and MMP2 Backbone'' <BR>
''DpnI digest of MMP2 insert and MMP2 Backbone'' <BR>
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After the PCR of the previous days I digested both the MMP2 insert and Backbone for the Gibson Assembly.
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-After the PCR of the previous days I digested both the MMP2 insert and Backbone for the Gibson Assembly.
''Gibson assembly for the MMP2+GFP construct'' <BR>
''Gibson assembly for the MMP2+GFP construct'' <BR>
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After the digest I did a Gibson assembly of the construct that contains the MMP2 gelatinase and superfolded GFP whose combined expression is induced by arabinose.
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-After the digest I did a Gibson assembly of the construct that contains the MMP2 gelatinase and superfolded GFP whose combined expression is induced by arabinose.
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''Transfromation of cells with the Cad-GFP Gibson Assembly Product (Sensing construct)'' <BR>
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''Transformation of cells with the Cad-GFP Gibson Assembly Product (Sensing construct)'' <BR>
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I transformed Cells with one cad-sensing Construct. They effectively grew, meaning that they did contain some sort of resistance. This indicated that the gibson had worked.
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-I transformed Cells with one cad-sensing Construct. They effectively grew, meaning that they did contain some sort of resistance. This indicated that the gibson had worked.
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Latest revision as of 22:17, 4 October 2013

Taxi.Coli: Smart Drug Delivery iGEM EPFL

Header

Cell Surface display

Characterization/Functional Assays
Inoculation of all our constructs and controls (competent cells, INP-Strepta-igem, INP-Strepta-dead, INP-Strepta-alive, BBa_K523013, YFP control, GFP control) needed for the confocal microscopy.

Sensing-Effector

PCR of MMP9 insert and GelE insert
-I did the PCR of the MMP9 and the gelE insert which worked. I then purified it which gave me concentrations around 100ng/ul.

DpnI digest of MMP2 insert and MMP2 Backbone
-After the PCR of the previous days I digested both the MMP2 insert and Backbone for the Gibson Assembly.

Gibson assembly for the MMP2+GFP construct
-After the digest I did a Gibson assembly of the construct that contains the MMP2 gelatinase and superfolded GFP whose combined expression is induced by arabinose.

Transformation of cells with the Cad-GFP Gibson Assembly Product (Sensing construct)
-I transformed Cells with one cad-sensing Construct. They effectively grew, meaning that they did contain some sort of resistance. This indicated that the gibson had worked.

Wiki Links

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