iGEM 2023 — Team ptsh-taiwan

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{% block title %}Notebook{% endblock %}
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# Notebook

* [March](#item-1)
* [April](#item-2)
* [May](#item-3)
* [June](#item-4)
* [July](#item-5)
* [August](#item-6)
* [September](#item-7)

## March

#### 3/4

* Topic decision
* Information collection

#### 3/11

* 7 chemical decided: Astaxanthin, NMN, PQQ, Vit B6, Butyrate, HMB(β-Hydroxy β-methylbutyric acid), KIC(α-Ketoisocaproic acid)
* The seven chemicals’ effects on human
* mTOR signal transduction pathway research

#### 3/18

* Market analyzing of the Sarcopenia
* Data collection on Sarcopenia
* Research on supportive treatment of Sarcopenia

![](https://static.igem.wiki/teams/4743/wiki/notebook-1.png)

#### 3/25

* Research on the cause of the Sarcopenia
* Research on the mechanism of Butyrate and HMB to boost muscle production
* Deeper research in mTOR pathway

![](https://static.igem.wiki/teams/4743/wiki/notebook-2.png)

## April

#### 4/1

* The connection of mTOR and Butyrate and HMB
* Research the mechanism of boosting muscle of other compounds:Astaxanthin, NMN, PQQ, Vit B6,and KIC
* Creating 31 pages report for our research

![](https://static.igem.wiki/teams/4743/wiki/notebook-3.png)![](https://static.igem.wiki/teams/4743/wiki/notebook-4.png)![](https://static.igem.wiki/teams/4743/wiki/notebook-5.png)

#### 4/8

* The connection of mTOR and Butyrate and HMB
* Research the mechanism of boosting muscle of other compounds:Astaxanthin, NMN, PQQ, Vit B6,and KIC
* Creating a 31 page report for our research on 7 chemical compounds

#### 4/15

* Decide the organism of producing these compounds
* Recruit new members
* Creating introductory presentation on PPT
* Done with our drawing of chemical compounds

![](https://static.igem.wiki/teams/4743/wiki/notebook-6.png)![](https://static.igem.wiki/teams/4743/wiki/notebook-7.png)

#### 4/22

* Distribute the works of producing chemical compounds to three probiotics
  1.Clostridium Butyricum: Butyrate, Vitamin B6
  2.Bacillus subtilis: PQQ, HMB, KIC
  3.Kluyveromyces marxianus: NMN, Astaxanthin
* Looking for metabolic pathways for producing these compounds on KEGG or other resource
* Done the First draft on metabolic pathway producing NMN

![](https://static.igem.wiki/teams/4743/wiki/notebook-8.png)

#### 4/29

* Decide the knockouts of the genes
* Starting to search for possible kill switch

![](https://static.igem.wiki/teams/4743/wiki/notebook-9.png)![](https://static.igem.wiki/teams/4743/wiki/notebook-10.png)

## May

#### 5/13

* Decide to concentrate on NMN,HMB,and KIC production
* Done the second draft of NMN producing pathway

![](https://static.igem.wiki/teams/4743/wiki/notebook-11.png)

#### 5/20

* Done the final draft of NMN producing pathway
* Designed our own pathway for NMN production

![](https://static.igem.wiki/teams/4743/wiki/notebook-12.png)![](https://static.igem.wiki/teams/4743/wiki/notebook-13.png)

#### 5/27

* Interview with doctors
* Kill switch design by using operon system
* Learned on biobrick build up

## June

#### 6/3

* Research on the conformation of the enzyme NAMPT(an enzyme essential for NMN)
* BLAST and search for the highest efficiency of NAMPT to produce most NMN
* Finalize the pathway to produce HMB and KIC

![](https://static.igem.wiki/teams/4743/wiki/notebook-14.png)

#### 6/10

* Keep researching on NAMPT
* Researching for the Intestine environment and the location of our probiotics will grow
* Researching for dosage of our chemical compounds

![](https://static.igem.wiki/teams/4743/wiki/notebook-15.png)![](https://static.igem.wiki/teams/4743/wiki/notebook-16.png)

#### 6/17

* Finding out sequence for what we want to insert and what we want to knock out in our metabolic pathway
* Design the second draft of our kill switch for Bacillus Subtilis

#### 6/24

* Finding out key active site of the NAMPT
* Design the third draft of our kill switch for Bacillus Subtilis
* Research for hydrogen sulfide sensitive repressor
* Finding Toxin and Antitoxin of Bacillus Subtilis

![](https://static.igem.wiki/teams/4743/wiki/notebook-17.png)![](https://static.igem.wiki/teams/4743/wiki/notebook-18.png)

## July

#### 7/1

* Draw out the kill switch for Bacillus Subtilis
* Drawing the first draft of kill switch of Clostridium and kluyveromyces

![](https://static.igem.wiki/teams/4743/wiki/notebook-19.png)

#### 7/8

* Choosing the expression vector of Bacillus subtilis
* Preparing for competent cells
* Draw out the gene map

![](https://static.igem.wiki/teams/4743/wiki/notebook-20.png)

#### 7/15

* Learning to use snapgene to draw map
* Design the map of kluyveromyces marxianus

![](https://static.igem.wiki/teams/4743/wiki/notebook-21.png)![](https://static.igem.wiki/teams/4743/wiki/notebook-22.png)

#### 7/22

* Send out the order of fragments
* Sequence optimization
* Optimize the design

#### 7/29

* Finalize the genes and order out
* Complete the transformation of TDH3,ADH1 promoter and GFP gene from the distribution kit
* Done the colon PCR

## August

#### 8/5

* Designing experiments

#### 8/12

* Start the experiments of NadA,NadB,and NadE fragments
* Ligation and transformation for the plasmid
* Gel extraction
* Check the length of pKlac2 plasmid

![](https://static.igem.wiki/teams/4743/wiki/notebook-23.png)![](https://static.igem.wiki/teams/4743/wiki/notebook-24.png)

#### 8/19

* Colony PCR checking
* Failed the ligation

![](https://static.igem.wiki/teams/4743/wiki/notebook-25.png)

#### 8/26

* Started Astaxanthin experiment
* Planning to redo the NMN experiment

## September

#### 9/2

* Wiki build up- HP and wet lab
* Final presentation making

#### 9/9

* Wiki build up- Projet,wet lab, and Human practice
* Enter register
* Started making promotion videos

#### 9/16

* Wiki build up- Projet,wet lab, and Human practice
* Checking for safety forms
* Finalize promotion videos

#### 9/23

* Wiki build up- Projet,wet lab, and Human practice
* Finalizing safety forms
* Finalize promotion videos

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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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#### Silver Medal Criterion #1

Demonstrate engineering success in a part of your project by going through at least one iteration of the engineering design cycle. This achievement should be distinct from your Contribution for Bronze.

If you plan to show engineering success by creating a new Part that has been shown to work as expected, you must document your contribution on the Part's Main Page on the [Registry](http://parts.igem.org/Main_Page) for your team to be eligible for this criteria.

---

Please see the [2023 Medals Page](https://competition.igem.org/judging/medals) for more information.

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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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## What should this page contain?

---

Describe the research, experiments, and protocols you used in your iGEM project. These should be detailed enough for another team to repeat your experiments.

If you made Parts this year, please remember to put all information, characterization, and measurement data on the Part's Main Page on the [Registry](http://parts.igem.org/Main_Page).

## Inspirations

---

* [2019 Nantes](https://2019.igem.org/Team%3ANantes/Experiments)
* [2019 TU Eindhoven](https://2019.igem.org/Team%3ATU_Eindhoven/Experiments)
* [2019 Mingdao](https://2019.igem.org/Team%3AMingdao/Demonstrate)
* [2020 Amsterdam](https://2020.igem.org/Team%3AAmsterdam/Experiments)
* [2020 NCTU Formosa](https://2020.igem.org/Team%3ANCTU_Formosa/Experiments)
* [2020 USAFA](https://2020.igem.org/Team%3AUSAFA/Experiments)

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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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## 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 also be on the Part's Main Page on the [Registry](http://parts.igem.org/Main_Page).** Otherwise these data will not be in consideration for any medals or part awards!
* 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.

## Inspirations

---

* [2019 Newcastle](https://2019.igem.org/Team%3ANewcastle/Results)
* [2019 Munich](https://2019.igem.org/Team%3AMunich/Results)
* [2019 Tec Chihuahua](https://2019.igem.org/Team%3ATec-Chihuahua/Results)
* [2020 Aalto Helsinki](https://2020.igem.org/Team%3AAalto-Helsinki/Results)
* [2020 GreatBay SCIE](https://2020.igem.org/Team%3AGreatBay_SCIE/Results)
* [2020 Queens Canada](https://2020.igem.org/Team%3AQueens_Canada/Results)

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