iGEM 2017 — Team Warwick

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# Team:Warwick/Notebook

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  + [Entrepreneurship](https://2017.igem.org/Team%3AWarwick/Entrepreneurship)
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  + [Software](https://2017.igem.org/Team%3AWarwick/Software)
* About
  + [Team](https://2017.igem.org/Team%3AWarwick/Team)
  + [Attributions](https://2017.igem.org/Team%3AWarwick/Attributions)
  + [Collaborations](https://2017.igem.org/Team%3AWarwick/Collaborations)

## Notebook

The PDF viewing plugin is not supported on your device, you may download the file [here](https://static.igem.org/mediawiki/2017/e/ee/T--Warwick--2-Notebook.pdf) instead.

We thank our sponsors without whom none of this would have been possible:

![](https://static.igem.org/mediawiki/2017/d/d0/T--Warwick--WISBlogo.png)

![](https://static.igem.org/mediawiki/2017/3/37/T--Warwick--SLSlogo.png)

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# Team:Warwick/Experiments

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* [Front page](https://2017.igem.org/Team%3AWarwick)
* Project
  + [Description](https://2017.igem.org/Team%3AWarwick/Description)
  + [Design](https://2017.igem.org/Team%3AWarwick/Design)
  + [Experiments](https://2017.igem.org/Team%3AWarwick/Experiments)
  + [Notebook](https://2017.igem.org/Team%3AWarwick/Notebook)
  + [Safety](https://2017.igem.org/Team%3AWarwick/Safety)
* Parts
  + [Parts](https://2017.igem.org/Team%3AWarwick/Basic_Part)
  + [Interlab](https://2017.igem.org/Team%3AWarwick/InterLab)
* Misc
  + [Public Engagement](https://2017.igem.org/Team%3AWarwick/Engagement)
  + [Silver Human Practices](https://2017.igem.org/Team%3AWarwick/HP/Silver)
  + [Gold Human Practices](https://2017.igem.org/Team%3AWarwick/HP/Gold_Integrated)
  + [Entrepreneurship](https://2017.igem.org/Team%3AWarwick/Entrepreneurship)
  + [Model](https://2017.igem.org/Team%3AWarwick/Model)
  + [Software](https://2017.igem.org/Team%3AWarwick/Software)
* About
  + [Team](https://2017.igem.org/Team%3AWarwick/Team)
  + [Attributions](https://2017.igem.org/Team%3AWarwick/Attributions)
  + [Collaborations](https://2017.igem.org/Team%3AWarwick/Collaborations)

## Experiments

The PDF viewing plugin is not supported on your device, you may download the file [here](https://static.igem.org/mediawiki/2017/8/85/T--Warwick--2-Completed-Lab-Book.pdf) instead.

Initially we began by splicing gene fragments provided by iGEM - due to the complexity of our part, we found it to be a lengthy process and time was running out. Professor John McCarthy suggested we use Gibson assembly, providing us space in his lab and assistance from his colleagues.

In late October when the fragments were due to arrive, preparations for the assembly revealed that the backbone the fragments had been prepared for was Kanamycin; however the iGEM registry only accept submissions from Chloramphenicol resistance marked backbones (pSB1C3 specifically).

![](https://static.igem.org/mediawiki/2017/a/a1/T--Warwick--glam5.png)

![](https://static.igem.org/mediawiki/2017/f/f5/T--Warwick--2-glam6.png)

In order to fix this, primers were prepared which would match the ends of the pSB1C3 linearised backbone. Tails on these primers had the same sequence as the tips of the Kanamycin backbone. By performing PCR, the pSB1C3 backbones could be modified in such a way to allow for Gibson assembly directly into the correct backbone.

![](https://static.igem.org/mediawiki/2017/a/ad/T--Warwick--2-Biobrick-Part-History.png)

### Three experiments had been considered for the modified cells:

A. Grow the cells on an IPTG gradient plate: Cells in greater concentration of IPTG should form thick biofilm, and cells in lower concentrations of IPTG will form thinner biofilms.

B. Grow the cells on an IPTG spot plate: Cells are to be spread over a plate and a drop of IPTG is to be placed and marked. Cells around the droplet of IPTG should form a thick biofilm, and cells in thinner areas will form less (E.G Part:BBa\_K777121 BBa\_K2012002)

C. Grow the cells on a lactose plate: A plate with Glucose, Lactose, and Arabanose is prepared. First the cells will degrade glucose - biofilm will be thin. Cells will then degrade Lactose - biofilm will become thicker. Cells will then degrade Arabanose - biofilm will become thin once more. By using a camera to perform a time-lapse, the Biofilm Manipulator's ability to control cellular adhesion and motility would be clearly defined.

*(Positive and negative control plates containing a constant IPTG concentration and no IPTG respectively should be used for comparison)*

We thank our sponsors without whom none of this would have been possible:

![](https://static.igem.org/mediawiki/2017/d/d0/T--Warwick--WISBlogo.png)

![](https://static.igem.org/mediawiki/2017/3/37/T--Warwick--SLSlogo.png)

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![](https://static.igem.org/mediawiki/2017/6/67/T--Warwick--BBSRClogo.png)

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# Team:Warwick/Results

# 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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