iGEM 2017 — Team NWU-CHINA

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# Team:NWU-CHINA/Notebook

* [HOME](https://2017.igem.org/Team%3ANWU-CHINA)
* [Team](https://2017.igem.org/Team%3ANWU-CHINA/Team)
* Project

  [Project-Home](https://2017.igem.org/Team%3ANWU-CHINA/Project-Home)

  ---

  [Preparation](https://2017.igem.org/Team%3ANWU-CHINA/Preparation)
  [Sense](https://2017.igem.org/Team%3ANWU-CHINA/Sense)
  [Expression](https://2017.igem.org/Team%3ANWU-CHINA/Expression)
  [Description](https://2017.igem.org/Team%3ANWU-CHINA/Description)
  [Notebook](https://2017.igem.org/Team%3ANWU-CHINA/Notebook)
  [Interlab](https://2017.igem.org/Team%3ANWU-CHINA/InterLab)
  [Attributions](https://2017.igem.org/Team%3ANWU-CHINA/Attributions)
* [Parts](https://2017.igem.org/Team%3ANWU-CHINA/Parts)
* [Safety](https://2017.igem.org/Team%3ANWU-CHINA/Safety)
* Human practice

  [Silver HP](https://2017.igem.org/Team%3ANWU-CHINA/HP/Silver)
  [Integrated HP](https://2017.igem.org/Team%3ANWU-CHINA/HP/Gold_Integrated)
  [Collaboration](https://2017.igem.org/Team%3ANWU-CHINA/Collaborations)
* [Achievement](https://2017.igem.org/Team%3ANWU-CHINA/Achievement)

![](https://static.igem.org/mediawiki/2017/5/55/NWU-CHINA-Project1.jpg)

# Notebook

March 15th- April 13th

We noticed Igem competition and get acquainted with synthesis biology.

 April 14th-May 1st

Our team, NWU-CHINA, had been organized. Then we find our PI and confirmed our project. Our PI gave us a lot of advice about following experiment.

 May 3rd-May 20th

We accepted foundational lab training and learned necessary experiment operation.

 June 1st-June 20th

We cleared Interlab Study requirement, RFC and 3A assembly protocol. Meanwhile, we found a lot of reference paper to find theory supporting our experiment.

 July 15th-July 26th

We finished our First Interlab Study

 August 1st-August 31st

Using Pak1900 to construct RFP vector and transferring constructed vector into P.a to verify expression of RFP in P.a .The detail protocol is as follows:

1. We got RFP sequence from registry and cloned RFP gene by PCR. Then we separated cloned RFP
2. gene by agarose gel electrophoresis and purified the gene by gel slices.
3. We used pAK1900 as our vector. Then we used BamH I and Sac I to digest pAK1900 and purified gene for 5 hours.
4. Then we used T4 DNA Ligase to ligated our vector and RFP gene at 4 ℃ for 24 hours
5. We transferred our vector into E.coli DH5&alpha competent cell to get more vector clones. Our transferring protocol is as follows:

1. Mix 4ul vector and 50ul competent cells in 1.5mL centrifuge tube at 0 ℃ for 30 min.
2. Hot shock for 90 seconds at 42℃ .
3. Keep competent cell at 0℃ for 4-5 min, then add 950&microL Soc media into the tube.
4. Incubate at 37 ℃ for 2 hours shaking at 220rpm
5. Centrifuge at 6000rpm for 1min. After centrifugation, 900&microL supernatant is discarded, then resuspended cells, spread the culture on plate with antibiotic 100&microg/&microL TC , incubate at 37℃ for 16 hours.
6. Select several colonies, purified them on another selective plate, incubate at 37℃ for 12 hours.

6. Incubate purified cultures with 5mL LB media and 5ul 5mg/&microL TC for 5 hours.
7. Using Plasmid minipreparation kit to extract the vector.

 September 1st-10th

We had done our second Interlab Study. Meanwhile, we used PCR to clone GntR and AlkB2 gene. And ligated them on constructed RFP vector. Then we separated cloned gene by agarose gel electrophoresis and purified the gene by gel slices.

 September 11th-October 12th

1. We used Hind I, Hind II, Sal I, Hind III and Xba I, Spe I and Sal I digest pAK1900, GntR, AlkB2 for 5 hours respectively.
2. Then we used T4 DNA Ligase to ligated them at 4℃ for 24 hours.
3. Then we cloned our vector in DH5&alpha competent cells.
4. Sequence our gene to verify our vector.

 October 12th-17th

We constructed our submission parts.

©2017 NWU-CHINA IGEM.All Rights Reserved.
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# Team:NWU-CHINA/Experiments

* [HOME](https://2017.igem.org/Team%3ANWU-CHINA)
* [Team](https://2017.igem.org/Team%3ANWU-CHINA/Team)
* Project

  [Project-Home](https://2017.igem.org/Team%3ANWU-CHINA/Project-Home)

  ---

  [Preparation](https://2017.igem.org/Team%3ANWU-CHINA/Preparation)
  [Sense](https://2017.igem.org/Team%3ANWU-CHINA/Sense)
  [Expression](https://2017.igem.org/Team%3ANWU-CHINA/Expression)
  [Description](https://2017.igem.org/Team%3ANWU-CHINA/Description)
  [Notebook](https://2017.igem.org/Team%3ANWU-CHINA/Notebook)
  [Interlab](https://2017.igem.org/Team%3ANWU-CHINA/InterLab)
  [Attributions](https://2017.igem.org/Team%3ANWU-CHINA/Attributions)
* [Parts](https://2017.igem.org/Team%3ANWU-CHINA/Parts)
* [Safety](https://2017.igem.org/Team%3ANWU-CHINA/Safety)
* Human practice

  [Silver HP](https://2017.igem.org/Team%3ANWU-CHINA/HP/Silver)
  [Integrated HP](https://2017.igem.org/Team%3ANWU-CHINA/HP/Gold_Integrated)
  [Collaboration](https://2017.igem.org/Team%3ANWU-CHINA/Collaborations)
* [Achievement](https://2017.igem.org/Team%3ANWU-CHINA/Achievement)

# 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)

[![](https://static.igem.org/mediawiki/2017/b/bf/HQ_menu_logo.jpg)](https://2017.igem.org)

[![](https://static.igem.org/mediawiki/2017/8/8b/HQ_page_logo.jpg)](https://2017.igem.org)

# Team:NWU-CHINA/Results

* [HOME](https://2017.igem.org/Team%3ANWU-CHINA)
* [Team](https://2017.igem.org/Team%3ANWU-CHINA/Team)
* Project

  [Project-Home](https://2017.igem.org/Team%3ANWU-CHINA/Project-Home)

  ---

  [Preparation](https://2017.igem.org/Team%3ANWU-CHINA/Preparation)
  [Sense](https://2017.igem.org/Team%3ANWU-CHINA/Sense)
  [Expression](https://2017.igem.org/Team%3ANWU-CHINA/Expression)
  [Description](https://2017.igem.org/Team%3ANWU-CHINA/Description)
  [Notebook](https://2017.igem.org/Team%3ANWU-CHINA/Notebook)
  [Interlab](https://2017.igem.org/Team%3ANWU-CHINA/InterLab)
  [Attributions](https://2017.igem.org/Team%3ANWU-CHINA/Attributions)
* [Parts](https://2017.igem.org/Team%3ANWU-CHINA/Parts)
* [Safety](https://2017.igem.org/Team%3ANWU-CHINA/Safety)
* Human practice

  [Silver HP](https://2017.igem.org/Team%3ANWU-CHINA/HP/Silver)
  [Integrated HP](https://2017.igem.org/Team%3ANWU-CHINA/HP/Gold_Integrated)
  [Collaboration](https://2017.igem.org/Team%3ANWU-CHINA/Collaborations)
* [Achievement](https://2017.igem.org/Team%3ANWU-CHINA/Achievement)

# 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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