Week 1: May 19 - May 26, 2013

We set up our labspace and started some preparatory work.

Tuesday, May 21, 2013

We finally moved in our lab. A bit of dust here and some rubbish to dispose there, but after a few hours of combined strength our lab was ready to go. Wet experiments can begin!

Thursday, May 23, 2013

Investigators: Kevin, Kerstin, Laura
We prepared some chemically competent XL1 blue E. Coli cells for all the transformations we are going to have to do during the project.

Friday, May 24, 2013

Investigators: Kevin, Kerstin, Laura
Today we transformed our freshly prepared competent cells for test purposes. We used the plasmids pUC 18 and pUC19 to calculate the transformation efficiency. 100 pg of DNA were used for each transformation with 50 µl cells. Additionally, competent cells were plated on ampicillin, kanamycin and chloramphenicol to ensure that the strain is not carrying any corresponding resistances. The plates were incubated at 37 °C over night.

Saturday, May 25, 2013

Investigators: Kevin, Kerstin, Laura
111 colonies were counted for pUC18 transformation, 50 for pUC19 transformation.
Transformation efficiency:
pUC18: ~ 10⁶ µg^-1
pUC19: ~5x10⁵ µg^-1
The plates with the additional antibiotics were empty, therefore the strain was not carrying any resistences.

Week 2: May 27 - June 1, 2013

The distribution kit arrived and we started with the actual labwork. 19 biobricks had to be transformed into our E. coli to secure the parts. Additionally we developed the cloning strategy for the next weeks.

Week 3: June 2 - June 8, 2013

This week we successfully cloned and transformed some of our first combination Bricks. We also managed to obtain some BioBricks that could not be transformed via Phusion-PCR directly from resuspended DNA.

Week 4: June 9 - June 15, 2013

This week we confirmed the successful cloning of the constitutive promoter + RBS. Furthermore we confirmed the addition of the TT to the autoinducer synthases and the lactonase. More biobrick sequences were verified. However, we observed some point mutations in the prefix/suffix sequences of some bricks as well as major annotation differences in the biobrick C0061.

Week 5: June 16 - June 22, 2013

This week was dominated by a lot of cloning. We equipped our constitutive and inducible promoters with ribosome binding sites, terminators, an ampicillin resistance gene and a lactonase (not all of them got each of these elements). Additionally, we separated the ampicillin resistance gene from its native promoter in order to make its expression inducible. Although cloning was mostly successful, we were struggling to amplify and purify the luxR brick (C0062), which cost us a lot of time and effort. Unfortunately, it also wasn’t rewarded with success in the end.

Another important step was to adapt our cloning strategy towards new chromoproteins which we received from iGEM team Uppsala. These allow us to evaluate our cultuvation experiments solely with own equipment, rather than having to find a cell sorter with suitable excitement lasers for all fluorescent proteins.

Week 6: June 23 - June 29, 2013

Week 6

Week 7: June 30 - July 6, 2013

In week 7 we started our first experiments with our inducible promotors. Unfortunatly we had trouble caused by the leakyness of two of our three promotors. We did further experiments but could not find a solution yet. Good news is that we received the chromoproteins from Uppsala! So we started working on our new cloning strategy by combining the chromoprotein DNA with our constitutive promotor-RBS construct. We hope to see nice and colorful colonies next week!

Week 8: July 7 - July 13, 2013

Week 8

Week 9: July 14 - July 20, 2013

Week 9

Week 10: July 21 - July 27, 2013

Week 10

Week 11: July 28 - August 3, 2013

Week 11

Week 12: August 4 - August 10, 2013

Week 11