iGEM 2018 — Team NPU-China

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# Team:NPU-China/Notebook

*menu*

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* + [HOME](https://2018.igem.org/Team%3ANPU-China)
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    - [Overview](https://2018.igem.org/Team%3ANPU-China/Overview)
    - [Background](https://2018.igem.org/Team%3ANPU-China/Background)
    - [Mini
      mtDNA Design](https://2018.igem.org/Team%3ANPU-China/Mini_mtDNA_Design)
    - [Synthesis](https://2018.igem.org/Team%3ANPU-China/Synthesis)
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      work](https://2018.igem.org/Team%3ANPU-China/Future_work)
  + Human Practices

    - [Intergrated](https://2018.igem.org/Team%3ANPU-China/Human_Practices)
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    - [Notebook](https://2018.igem.org/Team%3ANPU-China/Notebook)
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      Members](https://2018.igem.org/Team%3ANPU-China/Team_Members)
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  + For Judge

    - [Map](https://2018.igem.org/Team%3ANPU-China/Map)
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![](https://static.igem.org/mediawiki/2018/7/7d/T--NPU-China--bk-lab.png)

#### NoteBook

##### Chronological List of Progression

##### **Week 1** 15/07/2018 - 21/07/2018 Project Handover. Instruction of lab notices and experimental skills. Test remaining DNA fragments and re-prepare first-stage fragments with miscellaneous bands and degradation. Amplification of well preserved second-stage fragments. Confirm the skeleton and strain for homologous recombination in vivo: Y33-ura and By4741(S.c). Design a new series of primers especially for recombination in vivo: LL33-X-F/R. **Week 2** 22/07/2018 - 28/07/2018 Confirm the skeleton and strain for homologous recombination in vivo: Y33-ura and By4741(S.c). Design a new series of primers especially for recombination in vivo: LL33-X-F/R. Prepare the materials used for yeast transformation. Explore the PCR conditions for fragments with less amplification effect. Prepare new first-stage fragments with new series of primers: LLX. **Week3** 29/07/2018 - 04/08/2018 Prepare new first-stage fragments with new series of primers: LLX. Yeast transformation but failed. Find out the cause of failure and make it up. Prepare phosphorylated fragments(DX) for DATEL. Try DATEL and obtained much larger DNA fragments in vito. Determined to use new S.c strain: By4743 for transformation in vivo. **Week 4** 05/08/2018 - 11/08/2018 Design a new series of plasmids for yeast transformation verification Yeast transformation for Plasmid 1 and Plasmid 2 by using By4743. Some groups are verified successfully by verification primers. But encounter with obstacles during amplification of plasmids with in E.coli. Try to find out possible cause of failure. Continue DATEL but encounter with some obstacles. **Week 5** 12/08/2018 - 18/08/2018 Re-transformation in yeast for Plasmid 1 and Plasmid 2. Yeast transformation for Plasmid 3. Test transformation results. **Week 6** 19/08/2018 - 25/08/2018 Determine to use new skeleton stabilizing large geomic fragments: pGF. Re-construct the whole genome into 3 parts: All(All genomic fragments transformed in once), H1(Front part of genomic fragments transformed in once), and H2(Latter part of genomic fragments transformed in once). **Week 7** 26/08/2018 - 01/09/2018 Design new primers. Amplification of new fragments used for transformation. Explore the PCR conditions for new fragments. **Week 8** 02/09/2018 - 08/09/2018 Explore the PCR conditions for new fragments. Continue standardizing parts. **Week 9-10** 09/09/2018 - 22/09/2018 Prepare enough fragments for transformation. Debug failures during standardization. **Week 11** 23/09/2018 - 29/09/2018 Transformation in vivo for Plasmid ALL, Plasmid H1, Plasmid H2 but encounter with obstacles. Debug and find new way to solve them. **Week 12** 30/09/2018 - 06/10/2018 Transform again in vivo. Successfully obtain assembled Plasmid ALL and Plasmid H2 within yeast strain(H1 fail), after verification of verifying primers. **Week 13** 07/10/2018 - 13/10/2018 Try to amplify genomic fragments in E.coli( strain EPI300) by electrotransformation but failed. Successfully amplify fragment H2 (19kb) by PCR. Debug transformation(Too much anti-biotic), and re-transform successfully by chemical transformation. **Week 14** 14/10/2018 - Present Try to amplify Plasmid ALL by multicopy of pGF skeleton(OriV)

##### We have compiled an account of everything that has happened in the lab over the summer. We are excited to show you how busy we have been in iGEM 2018! We believe that sharing our work flow will convey the passion and dedication of iGEM students all around the world, as we continuously push our limits to achieve high quality results.

##### If you don't have a PDF plugin for this browser. No biggie... you can [click here to download the PDF file.](https://static.igem.org/mediawiki/2018/6/67/T--NPU-China--NoteBook.pdf)

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# Team:NPU-China/Experiments

# 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/2018/8/8b/HQ_page_logo.jpg)](https://2018.igem.org)

# Team:NPU-China/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.

### 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)