Team:EPF Lausanne/Calendar/7 September 2013

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(Created page with "{{Template:EPFL2013Header}} Sensing <BR> '''Purification of PCR products of the three sensing constructs''' <BR> I purified the PCR products from the previous Day and the conce...")
 
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<font size = "4"> Cell Surface Display </font> <BR>
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Research for the confocal microscopy.
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<br>Research of protocols for biotin fluorescent staining next for other functional assays.
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<br><br>
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Sensing <BR>
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<font size = "4"> Sensing-Effector </font> <BR>
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'''Purification of PCR products of the three sensing constructs''' <BR>
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''Purification of PCR products of the three sensing constructs'' <BR>
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I purified the PCR products from the previous Day and the concentrations were within an acceptable range. Since I had not yet amplified the constitutive promoter from iGEM I put the purified Backbone for this promoter into the refridgarator for late use.
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-I purified the PCR products from the previous Day and the concentrations were within an acceptable range. Since I had not yet amplified the constitutive promoter from iGEM I put the purified Backbone for this promoter into the refrigerator for late use.
 +
''DpnI digest of the two pH sensitive promoters and their backbones'' <BR>
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-I did a DpnI digest and measured the concentrations again, they were all around 50ng/ul which is good.
 +
''Gibson assembly of the two constructs with the hya and the cad promoter (pH-sensors)''<BR>
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-I did a Gibson assembly of both sensing constructs but it did not work for the one with the hya promoter, so I did another Restriction digest of the purified PCR products for this promoter.
-
'''DpnI digest of the two pH sensitive promoters and their backbones''' <BR>
+
''PCR of the MMP2 insert and its Backbone and the MMP9 Backbone'' <BR>
-
I did a DpnI digest and measured the concentrations again, they were all around 50ng/ul which is good.
+
-We did a PCR of the backbones for both MMP2 and MMP9 as well as of the insert MMP2. The Gel showed that it worked.
-
 
+
-
'''Gibson assembly of the two constructs with the hya and the cad promoter (pH-sensors) '''<BR>
+
-
I did a Gibson assembly of both sensning consruct but it did not work, so I did another Restriction digest of the purified PCR products.
+
-
 
+
-
'''PCR of the MMP2 insert and its Backbone and the MMP9 Backbone''' <BR>
+
-
We did a PCR of the backbones for both MMP2 and MMP9 as well as of the insertMMP2. The Gel showed that it worked.
+
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{{Template:EPFL2013Footer}}

Latest revision as of 22:16, 4 October 2013

Taxi.Coli: Smart Drug Delivery iGEM EPFL

Header

Cell Surface Display
Research for the confocal microscopy.
Research of protocols for biotin fluorescent staining next for other functional assays.

Sensing-Effector

Purification of PCR products of the three sensing constructs
-I purified the PCR products from the previous Day and the concentrations were within an acceptable range. Since I had not yet amplified the constitutive promoter from iGEM I put the purified Backbone for this promoter into the refrigerator for late use.

DpnI digest of the two pH sensitive promoters and their backbones
-I did a DpnI digest and measured the concentrations again, they were all around 50ng/ul which is good.

Gibson assembly of the two constructs with the hya and the cad promoter (pH-sensors)
-I did a Gibson assembly of both sensing constructs but it did not work for the one with the hya promoter, so I did another Restriction digest of the purified PCR products for this promoter.

PCR of the MMP2 insert and its Backbone and the MMP9 Backbone
-We did a PCR of the backbones for both MMP2 and MMP9 as well as of the insert MMP2. The Gel showed that it worked.

Wiki Links

Retrieved from "http://2013.igem.org/Team:EPF_Lausanne/Calendar/7_September_2013"