iGEM 2015 — Team TU_Eindhoven

# Team:TU Eindhoven/Notebook

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Safety](https://2015.igem.org/Team%3ATU_Eindhoven/Safety)

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[Team members](https://2015.igem.org/Team%3ATU_Eindhoven/Team/Members)[Supervisors](https://2015.igem.org/Team%3ATU_Eindhoven/Team/Supervisors)[Attributions](https://2015.igem.org/Team%3ATU_Eindhoven/Attributions)

[Description](https://2015.igem.org/Team%3ATU_Eindhoven/Description)[Design](https://2015.igem.org/Team%3ATU_Eindhoven/Project/Design)[Expression System](https://2015.igem.org/Team%3ATU_Eindhoven/Project/Expression_System)[Experimental Approach](https://2015.igem.org/Team%3ATU_Eindhoven/Project/Experimental_Approach)[Protocols](https://2015.igem.org/Team%3ATU_Eindhoven/Project/Protocols)[Timeline](https://2015.igem.org/Team%3ATU_Eindhoven/Notebook)

[Parts](https://2015.igem.org/Team%3ATU_Eindhoven/Parts)[Results](https://2015.igem.org/Team%3ATU_Eindhoven/Results)[Medal Motivation](https://2015.igem.org/Team%3ATU_Eindhoven/Achievements)[Interlab Study](https://2015.igem.org/Team%3ATU_Eindhoven/Measurement)

[Approach](https://2015.igem.org/Team%3ATU_Eindhoven/Modeling/Approach)[Script](https://2015.igem.org/Team%3ATU_Eindhoven/Modeling/Script)[Results](https://2015.igem.org/Team%3ATU_Eindhoven/Modeling/Results)

[Outreach](https://2015.igem.org/Team%3ATU_Eindhoven/Policy_Practices/Human_Outreach)[Application Scenarios](https://2015.igem.org/Team%3ATU_Eindhoven/Policy_Practices/Application_scenarios)[Stakeholders](https://2015.igem.org/Team%3ATU_Eindhoven/Policy_Practices/Stakeholders)[Regulations](https://2015.igem.org/Team%3ATU_Eindhoven/Policy_Practices/Regulations)

[Cloning Guide](https://2015.igem.org/Team%3ATU_Eindhoven/Collaborations/CloningGuide)[Collaboration Attributions](https://2015.igem.org/Team%3ATU_Eindhoven/Collaborations/Collaboration_Attributions)

[Lab Safety](https://2015.igem.org/Team%3ATU_Eindhoven/Safety/Lab)[Project Safety](https://2015.igem.org/Team%3ATU_Eindhoven/Safety/Project)[Lab Equipment](https://2015.igem.org/Team%3ATU_Eindhoven/Safety/LabEquipment)

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Dig Even Deeper
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* See our [Protocols](https://2015.igem.org/Team%3ATU_Eindhoven/Project/Protocols)

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See how modelling helped us understand our device](https://2015.igem.org/Team%3ATU_Eindhoven/Modeling)
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Missed something? Jump back to our Project page](https://2015.igem.org/Team%3ATU_Eindhoven/Project)

# Timeline

Somewhere in the beginning of July, we started our wetwork. We started the summer off working in smaller teams: we had a team working on Gibson Assembly, a team responsible for Traditional Cloning and a team working on Interlab. In this way, we could fit tons of wetwork within the few weeks of the summer holidays. Below, we hope to give a short overview of how we started off and which experiments we carried out in which weeks.

See the black timeline below for a global impression of the major experiments carried out by the teams working with Gibson Assembly and Traditional cloning. These experiments brougth us were we are today and results can be seen [here](https://2015.igem.org/Team%3ATU_Eindhoven/ResultsHome).

#### Week 26

### - Preparing for take-off

![](https://static.igem.org/mediawiki/2015/8/87/TU_Eindhoven_Ingeklapt.png)

* Inventory of the lab supplies
* Pouring LB Agar plates
* Amplification of the pET-Duet-1 vector
* Assessment of the safety requirements
* Preparing stocks for antibiotics, glycerol, LB & MilliQ

#### Week 27

### - The clone wars

![](https://static.igem.org/mediawiki/2015/8/87/TU_Eindhoven_Ingeklapt.png)

Gibson Assembly:

* Linearizing MCS-1 of pET-Duet-1 for Gibson Assembly
* Digestion of the template using DpnI
* Running a gel to check whether the linearization was successful
* Our first Gibson Assembly!
* Plasmid amplification into NEB 5-alpha

 Traditional cloning:

* Amplification of the pET-Duet-1 vector
* Nde1 & Kpn1 digestion of the pET-Duet-1 vector (MCS-2) for traditional cloning

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