iGEM 2017 — Team Tuebingen

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# Team:Tuebingen/Lab/Notebook

iGEM Tübingen 2017

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[Team](https://2017.igem.org/Team%3ATuebingen/Team)
[Inspiration](https://2017.igem.org/Team%3ATuebingen/Inspiration)
[Results](https://2017.igem.org/Team%3ATuebingen/Demonstrate)
[Human Practice](https://2017.igem.org/Team%3ATuebingen/HP)
[Lab](https://2017.igem.org/Team%3ATuebingen/Lab)
[Attributions](https://2017.igem.org/Team%3ATuebingen/Attributions)

![InterLabBild](https://static.igem.org/mediawiki/2017/8/8c/T--Tuebingen--LabTitle.jpg)

lab book

[May 2017](#May 2017)
[June 2017](#June 2017)
[July 2017](#July 2017)
[August 2017](#August 2017)
[September 2017](#September 2017)
[October 2017](#October 2017)

## May 2017

* Begin of 3-(((1-carboxypropan-2-yl)amino)methyl)-4-hydroxybenzoic acid synthesis in a small batch.

## June 2017

* Duff-reaction and reductive amination occurred without problems.
* Synthesis of 3-(((1-carboxypropan-2-yl)amino)methyl)-4-hydroxybenzoic acid in a large batch succeeded without problems. Various cleanup methods were tried but failed.

## July 2017

* Various clean-up methods (extraction, recrystallization, chromatography) of chemical synthesis were tried, but failed. The product was stored in methanol at room temperature during this time, which might have reduced yield significantly.
* Preparation for work in molecular biology lab (competent cells, media, agar-plates). Begin of cloning β-lactam-synthetases wild type (WT) and double mutant (DM) into pUWL.

## August 2017

* No success in purification of 3-(((1-carboxypropan-2-yl)amino)methyl)-4-hydroxybenzoic acid.
* Begin of cloning NovL, SimL, CouL and CloL into pUWL and pSB1C3.
* The cloning of GyrB (*S.aureus*) plus Terminator BamHI/HindIII in pSB1C3 was started and finished successfully. Furthermore the GyrB (*E.coli*) was cloned into pSB4C5 under control of a arac-pBad promoter.
* Additionally the Interlabstudy was carried out.
* The work with *S. coelicolor* was started. Collection of spores from *S. coelicolor* M1146/Clo BG1 and Clo SA02 and inoculation of *S. coelicolor* M1146 / Clo BG1 for Clorobiocin production.

## September 2017

* Extraction of Clorobiocin from *S. coelicolor* M1146 / Clo BG1. HPLC analysis showed very low yield which made further purification not feasible.
* Cloning of CouL and CloL in pUWL was finished and conjugated into *S. coelicolor* M1146/SA02.
* Cloning of SimL and NovL did not work and was stopped to focus on more important projects.
* Cloning of BLS-WT in pSB1C3 was finished and confirmed by sequencing.
* First feeding experiments of *S. coelicolor* M1156/Clo BG1, with unpurified 3-(((1-carboxypropan-2-yl)amino)methyl)-4-hydroxybenzoic acid was started. LC-MS showed neither product nor intermediate.

## October 2017

* Due to the short time that was left, further feeding experiments were done with unpurified 3-(((1-carboxypropan-2-yl)amino)methyl)-4-hydroxybenzoic acid.
* Cloning of pRha (BBa\_K914003) and RBS (BBa\_B0034) into pSB1C3.
* Cloning of GyrB (*E.coli*) under control of an arac-pBad promoter into pSB1C3-pRha-RBS.
* Reordering of SHV and GyrA (*S. coelicolor*) gBlocks and cloning into pSB1C3. SHV cloning into pSB1C3 and pSB1C3-pRha-RBS worked immediately.
* Cloning of BLS DM in pSB1C3 and BLS WT in pSB1C3-pRha-RBS.
* Cloning of CloL and CouL into pSB1C3.
* Since cloning of BLS DM in pSB1C3 pRha-RBS did not work, it was cloned into pRha-RBS-pSB1K3 for test expression.
* Agar diffusion test with different concentrations of Clorobiocin with XL-1 blue and TolC *E.coli* were done. Clorobiocin and wafers were sent to iGEM team Franconia for our collaboration.
* Test expression and purification of BLS WT and DM.
* First try to close the ring of our chemical synthesis *in vitro*.

![](https://static.igem.org/mediawiki/2017/5/58/T%C3%BCbingen_Qiagen-Logo.jpeg) ![](https://static.igem.org/mediawiki/2017/1/1c/T%C3%BCbingen_Roth-Logo.jpeg) ![](https://static.igem.org/mediawiki/2017/d/d8/TuebingenZymoLogo.jpeg) ![](https://static.igem.org/mediawiki/2017/5/5b/T%C3%BCbingen_Eurofins-Logo.png) ![](https://static.igem.org/mediawiki/2017/e/e4/GATCTuebingen.png) ![](https://static.igem.org/mediawiki/2017/4/49/T%C3%BCbingen_NEB-Logo.jpeg)
![](https://static.igem.org/mediawiki/2017/7/7c/T%C3%BCbingen_Greiner-Logo.jpeg) ![](https://static.igem.org/mediawiki/2017/b/bf/T%C3%BCbingen_Agilent-Logo.jpeg) ![](https://static.igem.org/mediawiki/2017/0/01/T%C3%BCbingen_abcr-Logo.png) ![](https://static.igem.org/mediawiki/2017/5/59/MicrosynthTuebingen.png) ![](https://static.igem.org/mediawiki/2017/8/8b/Eppendorftuebingen.png)

© iGEM Team Tuebingen 2017

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# Team:Tuebingen/Experiments

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MENU

[HOME](https://2017.igem.org/Team%3ATuebingen)

TEAM

[Team](https://2017.igem.org/Team%3ATuebingen/Team)
[Collaborations](https://2017.igem.org/Team%3ATuebingen/Collaborations)

PROJECT

[Description](https://2017.igem.org/Team%3ATuebingen/Description)
[Design](https://2017.igem.org/Team%3ATuebingen/Design)
[Experiments](https://2017.igem.org/Team%3ATuebingen/Experiments)
[Notebook](https://2017.igem.org/Team%3ATuebingen/Notebook)
[InterLab](https://2017.igem.org/Team%3ATuebingen/InterLab)
[Contribution](https://2017.igem.org/Team%3ATuebingen/Contribution)
[Model](https://2017.igem.org/Team%3ATuebingen/Model)
[Results](https://2017.igem.org/Team%3ATuebingen/Results)
[Demonstrate](https://2017.igem.org/Team%3ATuebingen/Demonstrate)
[Improve](https://2017.igem.org/Team%3ATuebingen/Improve)
[Attributions](https://2017.igem.org/Team%3ATuebingen/Attributions)

PARTS

[Parts](https://2017.igem.org/Team%3ATuebingen/Parts)
[Basic Parts](https://2017.igem.org/Team%3ATuebingen/Basic_Part)
[Composite Parts](https://2017.igem.org/Team%3ATuebingen/Composite_Part)
[Part Collection](https://2017.igem.org/Team%3ATuebingen/Part_Collection)

[SAFETY](https://2017.igem.org/Team%3ATuebingen/Safety)

HUMAN PRACTICES

[Silver HP](https://2017.igem.org/Team%3ATuebingen/HP/Silver)
[Integrated and Gold](https://2017.igem.org/Team%3ATuebingen/HP/Gold_Integrated)
[Public Engagement](https://2017.igem.org/Team%3ATuebingen/Engagement)

AWARDS

[Applied Design](https://2017.igem.org/Team%3ATuebingen/Applied_Design)
[Entrepreneurship](https://2017.igem.org/Team%3ATuebingen/Entrepreneurship)
[Hardware](https://2017.igem.org/Team%3ATuebingen/Hardware)
[Measurement](https://2017.igem.org/Team%3ATuebingen/Measurement)
[Model](https://2017.igem.org/Team%3ATuebingen/Model)
[Plant](https://2017.igem.org/Team%3ATuebingen/Plant)
[Software](https://2017.igem.org/Team%3ATuebingen/Software)

[JUDGING FORM](https://igem.org/2017_Judging_Form?team=Tuebingen)

# Tuebingen

# Experiments

Describe the research, experiments, and protocols you used in your iGEM project. These should be detailed enough for another team to repeat your experiments.

Please remember to put all characterization and measurement data for your parts on the corresponding Registry part pages.

##### What should this page contain?

* Protocols
* Experiments
* Documentation of the development of your project

##### Inspiration

* [2014 Colombia](https://2014.igem.org/Team%3AColombia/Protocols)
* [2014 Imperial](https://2014.igem.org/Team%3AImperial/Protocols)
* [2014 Caltech](https://2014.igem.org/Team%3ACaltech/Project/Experiments)

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# Team:Tuebingen/Results

[![](http://placehold.it/350x150)](https://2017.igem.org/Team%3ATuebingen)

MENU

[HOME](https://2017.igem.org/Team%3ATuebingen)

TEAM

[Team](https://2017.igem.org/Team%3ATuebingen/Team)
[Collaborations](https://2017.igem.org/Team%3ATuebingen/Collaborations)

PROJECT

[Description](https://2017.igem.org/Team%3ATuebingen/Description)
[Design](https://2017.igem.org/Team%3ATuebingen/Design)
[Experiments](https://2017.igem.org/Team%3ATuebingen/Experiments)
[Notebook](https://2017.igem.org/Team%3ATuebingen/Notebook)
[InterLab](https://2017.igem.org/Team%3ATuebingen/InterLab)
[Contribution](https://2017.igem.org/Team%3ATuebingen/Contribution)
[Model](https://2017.igem.org/Team%3ATuebingen/Model)
[Results](https://2017.igem.org/Team%3ATuebingen/Results)
[Demonstrate](https://2017.igem.org/Team%3ATuebingen/Demonstrate)
[Improve](https://2017.igem.org/Team%3ATuebingen/Improve)
[Attributions](https://2017.igem.org/Team%3ATuebingen/Attributions)

PARTS

[Parts](https://2017.igem.org/Team%3ATuebingen/Parts)
[Basic Parts](https://2017.igem.org/Team%3ATuebingen/Basic_Part)
[Composite Parts](https://2017.igem.org/Team%3ATuebingen/Composite_Part)
[Part Collection](https://2017.igem.org/Team%3ATuebingen/Part_Collection)

[SAFETY](https://2017.igem.org/Team%3ATuebingen/Safety)

HUMAN PRACTICES

[Silver HP](https://2017.igem.org/Team%3ATuebingen/HP/Silver)
[Integrated and Gold](https://2017.igem.org/Team%3ATuebingen/HP/Gold_Integrated)
[Public Engagement](https://2017.igem.org/Team%3ATuebingen/Engagement)

AWARDS

[Applied Design](https://2017.igem.org/Team%3ATuebingen/Applied_Design)
[Entrepreneurship](https://2017.igem.org/Team%3ATuebingen/Entrepreneurship)
[Hardware](https://2017.igem.org/Team%3ATuebingen/Hardware)
[Measurement](https://2017.igem.org/Team%3ATuebingen/Measurement)
[Model](https://2017.igem.org/Team%3ATuebingen/Model)
[Plant](https://2017.igem.org/Team%3ATuebingen/Plant)
[Software](https://2017.igem.org/Team%3ATuebingen/Software)

[JUDGING FORM](https://igem.org/2017_Judging_Form?team=Tuebingen)

# Tuebingen

# Results

Here you can describe the results of your project and your future plans.

##### What should this page contain?

* Clearly and objectively describe the results of your work.
* Future plans for the project.
* Considerations for replicating the experiments.

##### You should also describe what your results mean:

* Interpretation of the results obtained during your project. Don't just show a plot/figure/graph/other, tell us what you think the data means. This is an important part of your project that the judges will look for.
* Show data, but remember all measurement and characterization data must be on part pages in the Registry.
* Consider including an analysis summary section to discuss what your results mean. Judges like to read what you think your data means, beyond all the data you have acquired during your project.

##### Project Achievements

You can also include a list of bullet points (and links) of the successes and failures you have had over your summer. It is a quick reference page for the judges to see what you achieved during your summer.

* A list of linked bullet points of the successful results during your project
* A list of linked bullet points of the unsuccessful results during your project. This is about being scientifically honest. If you worked on an area for a long time with no success, tell us so we know where you put your effort.

##### Inspiration

See how other teams presented their results.

* [2014 TU Darmstadt](https://2014.igem.org/Team%3ATU_Darmstadt/Results/Pathway)
* [2014 Imperial](https://2014.igem.org/Team%3AImperial/Results)
* [2014 Paris Bettencourt](https://2014.igem.org/Team%3AParis_Bettencourt/Results)

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