iGEM 2022 — Team hbut-china

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{% block title %}Notebook{% endblock %}
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![](https://static.igem.wiki/teams/4249/wiki/notebook/notebook-material-00.png)

![](https://static.igem.wiki/teams/4249/wiki/parts/parts-img-02.png)

| Date | Events |
| --- | --- |
|  | Brainstorming（Topic design, literature investigation） |
|  | Design and synthesis of gene sequences and primers |
| [3.24-3.30](https://static.igem.wiki/teams/4249/wiki/implementation-docs/3-24-3-30.pdf) | Double digest pML104, sgRNA hybridized, T4 ligation |
| [3.31-4.03](https://static.igem.wiki/teams/4249/wiki/implementation-docs/3-31-4-3.pdf) | Transformation and verification of recombination plasmid |
| [4.04-5.09](https://static.igem.wiki/teams/4249/wiki/implementation-docs/4-4-5-9.pdf) | Construction of recombination plasmid pUC57-pADH1-NcEgt1-tCYC1 |
| Construction of recombination plasmid pUC57-H1-pADH1-NcEgt1-tCYC1-H2 |
| [5.10-5.14](https://static.igem.wiki/teams/4249/wiki/implementation-docs/5-10-5-14.pdf) | Transformation with donor DNA and recombination plasmid |
| [5.15-5.21](https://static.igem.wiki/teams/4249/wiki/implementation-docs/5-15-5-21.pdf) | Selection of positive transformants and loss of sgRNA/Cas9 expression plasmid |
| [5.22-5.28](https://static.igem.wiki/teams/4249/wiki/implementation-docs/5-22-5-28.pdf) | Fermentation culture of recombinant strain and detection of target product by HPLC |
| 5.29-6.03 | Detection of target product by LC-MS |
| [6.04-6.16](https://static.igem.wiki/teams/4249/wiki/implementation-docs/6-4-6-16.pdf) | Construction of recombination plasmid pTEF2-pTEF1-CpEgt2-tCYC1 |
| Construction of recombination plasmid pTEF2-H1-pTEF1-CpEgt2-tCYC1-H2 |
| [6.17-6.22](https://static.igem.wiki/teams/4249/wiki/implementation-docs/6-17-6-22.pdf) | Construction of recombination pML104 targeting Saccharomyces cerevisiae chromosome 11 |
| [6.23-7.04](https://static.igem.wiki/teams/4249/wiki/implementation-docs/6-23-7-4.pdf) | Transformation with donor DNA and recombination plasmid |
| [7.05-7.10](https://static.igem.wiki/teams/4249/wiki/implementation-docs/7-5-7-10.pdf) | Selection of positive transformants and loss of sgRNA/Cas9 expression plasmid |
| [7.11-7.22](https://static.igem.wiki/teams/4249/wiki/implementation-docs/7-11-7-22.pdf) | Testing of EGT production capacity of two engineered strains |
| [7.23-8.02](https://static.igem.wiki/teams/4249/wiki/implementation-docs/7-23-8-2.pdf) | Determination of growth curve |

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{% block title %}Engineering Success{% endblock %}
{% block lead %}Demonstrate engineering success in a part of your project by going through at least one iteration of the
engineering design cycle.{% endblock %}
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![](https://static.igem.wiki/teams/4249/wiki/engineering/engineering-material-00.png)

![](https://static.igem.wiki/teams/4249/wiki/engineering/engineering-material-01.png)

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{% block title %}Experiments{% endblock %}
{% block lead %}Describe the research, experiments, and protocols you used in your iGEM project.{% endblock %}
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![](https://static.igem.wiki/teams/4249/wiki/experiments/experiments-material-00.png)

![](https://static.igem.wiki/teams/4249/wiki/engineering/engineering-material-01.png)

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{% block title %}Results{% endblock %}
{% block lead %}You can describe the results of your project and your future plans here.{% endblock %}
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![](https://static.igem.wiki/teams/4249/wiki/results/results-material-00.png)

![](https://static.igem.wiki/teams/4249/wiki/results/results-material-01.png)

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