Team:EPF Lausanne/Calendar/25 October 2013

From 2013.igem.org

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<br>- For ISY: PCR (6) to add Gibson overhangs on BBa_K523013_PCR_product (4) where linker were added the day before
<br>- For ISY: PCR (6) to add Gibson overhangs on BBa_K523013_PCR_product (4) where linker were added the day before
<br>- PCR purification
<br>- PCR purification
-
 
+
<br><br>
2) Gibson Assembly
2) Gibson Assembly
<br>- IYS Gibson Assembly using PCR (1) from the day before and PCR (5)
<br>- IYS Gibson Assembly using PCR (1) from the day before and PCR (5)
<br>- ISY Gibson Assembly using PCR (2) from the day before and PCR (6)
<br>- ISY Gibson Assembly using PCR (2) from the day before and PCR (6)
-
<b>
+
<br>- 0.8% Gel electrophoresis to verify the thirs PCRs and the Gibson Products => the products are higher than the expected sizes but we decided to do the transformation anyway.
-
<br>- 0.8% Gel electrophoresis to verify the thirs PCRs and the Gibson Products
+
<br><br>
<br><br>
 +
2)DH5_alpha Transformation
 +
<br>- Transformation of the two Gibson products with 2 controls consisting of the backbone and the insert use for the Gibson (PCR (1) & (5) for IYS and PCR (2) & (6) for ISY)
 +
<br>- Additional controls => competent cells and BBa_K523013 miniprep (initial template used for the PCRs)
 +
<br>- cells were plated on Chloramphenicol plates and grow overnight in a 37°C incubator
 +
<br><br>
 +
 +
<font size = "4"> Nanoparticles </font> <BR>
 +
 +
- Microfluidics experiment for nanoparticles digestion with MMP2
 +
 +
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Latest revision as of 22:10, 27 October 2013

Taxi.Coli: Smart Drug Delivery iGEM EPFL

Header

Cell Surface Display
CLoning INP_EYFP_Streptavidin (IYS) and INP_Streptavidin_EYFP (ISY)

1) Third PCRs (50µl)
- For IYS: PCR (5) to add Gibson overhangs on BBa_K523013_PCR_product (3) where linker were added the day before
- For ISY: PCR (6) to add Gibson overhangs on BBa_K523013_PCR_product (4) where linker were added the day before
- PCR purification

2) Gibson Assembly
- IYS Gibson Assembly using PCR (1) from the day before and PCR (5)
- ISY Gibson Assembly using PCR (2) from the day before and PCR (6)


- 0.8% Gel electrophoresis to verify the thirs PCRs and the Gibson Products => the products are higher than the expected sizes but we decided to do the transformation anyway.

2)DH5_alpha Transformation
- Transformation of the two Gibson products with 2 controls consisting of the backbone and the insert use for the Gibson (PCR (1) & (5) for IYS and PCR (2) & (6) for ISY)
- Additional controls => competent cells and BBa_K523013 miniprep (initial template used for the PCRs)
- cells were plated on Chloramphenicol plates and grow overnight in a 37°C incubator

Nanoparticles

- Microfluidics experiment for nanoparticles digestion with MMP2