Team:KULeuven/17 July 2008

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=== Wet Lab ===
=== Wet Lab ===
-
* The cells we transformed yesterday (TOP10 cells with BBa_30109 and DH5alpha with BBa_0034) did not grow on the ampicillin plate. The pUC control did grow on both plates, but the efficiency of the transformation with TOP10 cells seems to be better.
+
The cells we transformed yesterday (TOP10 cells with [http://partsregistry.org/Part:BBa_M30109 M30109] and DH5alpha with [http://partsregistry.org/Part:BBa_B0034 B0034]) did not grow on the ampicillin plates. The pUC control did grow on both plates, but the efficiency of the transformation with TOP10 cells seems to be better. There was a second attempt to transform TOP10 cells using the heat shock method from iGEM with some minor changes.
-
* Today we tried to transform TOP10 cells with the electroporation method. We used:
+
 
-
** plasmid with BBa_B0034
+
-
** plasmid with BBa_J6110 (this is a part we don't need for our project, so we don't lose any DNA during our attempts to transform cells)
+
-
** plasmid with BBa_J6110 (2 punches in 5ul TE-buffer)
+
-
** pUC control plasmid
+
-
* There was a second attempt to transform TOP10 cells using the heat shock method from iGEM with some minor changes.
+
-
* The protocol to make competent cells was continued. The grown TOP10 cells were inoculated in SOB broth and incubated for 8h. Then we inoculated them in a bigger volume SOB broth and let them incubate overnight.
+
Today one of our engineers, Maarten, set his first steps in the lab. We taught him how to pipette!
Today one of our engineers, Maarten, set his first steps in the lab. We taught him how to pipette!
 +
 +
We continued the protocol to make competent cells. The grown TOP10 cells were inoculated in SOB broth and incubated at 37°C for 8h. Then we inoculated them in a bigger volume SOB broth and let them incubate overnight.
 +
 +
Finally, we tried to transform TOP10 cells with the electroporation method. We used the following parts:
 +
* plasmid with [http://partsregistry.org/Part:BBa_B0034 B0034]
 +
* plasmid with [http://partsregistry.org/Part:BBa_J61100 J61100] (this is a part we don't need for our project, so we don't lose any DNA during our attempts to transform cells)
 +
* plasmid with [http://partsregistry.org/Part:BBa_J61100 J61100] (2 punches in 5ul TE-buffer in order to obtain a higher concentration of DNA)
 +
* pUC control plasmid
=== Dry Lab ===
=== Dry Lab ===
-
== Modeling ==
+
==== Modeling ====
-
Even more parameters ... Most composite parts are now realistically parametrized. Only the memory still remains and pieces of the pulse generator. This will be food for tomorrow.
+
Even more parameters - most composite parts are now realistically parameterized. Only the memory and pieces of the pulse generator remain. This will be food for tomorrow. The ''Pulse Generator'' was modeled, eventhough there are a few missing parameters left, but it does his job.
-
We have also started to put the composite parts together. Results, pictures and models (yay!) can be found in the modeling section. Things are actually looking surprisingly good :)
+
-
== Remarks ==
+
We have also started to put the composite parts together. Results, pictures and models (yay!) can be found in the ''modeling'' section. Things are actually looking surprisingly good :)

Latest revision as of 18:06, 6 October 2008

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Contents

Lab Work

Wet Lab

The cells we transformed yesterday (TOP10 cells with [http://partsregistry.org/Part:BBa_M30109 M30109] and DH5alpha with [http://partsregistry.org/Part:BBa_B0034 B0034]) did not grow on the ampicillin plates. The pUC control did grow on both plates, but the efficiency of the transformation with TOP10 cells seems to be better. There was a second attempt to transform TOP10 cells using the heat shock method from iGEM with some minor changes.

Today one of our engineers, Maarten, set his first steps in the lab. We taught him how to pipette!

We continued the protocol to make competent cells. The grown TOP10 cells were inoculated in SOB broth and incubated at 37°C for 8h. Then we inoculated them in a bigger volume SOB broth and let them incubate overnight.

Finally, we tried to transform TOP10 cells with the electroporation method. We used the following parts:

  • plasmid with [http://partsregistry.org/Part:BBa_B0034 B0034]
  • plasmid with [http://partsregistry.org/Part:BBa_J61100 J61100] (this is a part we don't need for our project, so we don't lose any DNA during our attempts to transform cells)
  • plasmid with [http://partsregistry.org/Part:BBa_J61100 J61100] (2 punches in 5ul TE-buffer in order to obtain a higher concentration of DNA)
  • pUC control plasmid

Dry Lab

Modeling

Even more parameters - most composite parts are now realistically parameterized. Only the memory and pieces of the pulse generator remain. This will be food for tomorrow. The Pulse Generator was modeled, eventhough there are a few missing parameters left, but it does his job.

We have also started to put the composite parts together. Results, pictures and models (yay!) can be found in the modeling section. Things are actually looking surprisingly good :)

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