Team:EPF Lausanne/Calendar/7 September 2013

From 2013.igem.org

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<font size = "4"> Sensing </font> <BR>
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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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<font size = "4"> Sensing-Effector </font> <BR>
''Purification of PCR products of the three sensing constructs'' <BR>
''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>
''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.
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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>
''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 construcst 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.
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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.
''PCR of the MMP2 insert and its Backbone and the MMP9 Backbone'' <BR>
''PCR of the MMP2 insert and its Backbone and the MMP9 Backbone'' <BR>
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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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-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.
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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.