iGEM 2020 — Team OSA

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:OSA/Notebook

# Notebook

Document the dates you worked on your project. This should be a detailed account of the work done each day for your project.

### What should this page have?

* Chronological notes of what your team is doing.
* Brief descriptions of daily important events.
* Pictures of your progress.
* Mention who participated in what task.

### Inspiration

You can see what others teams have done to organize their notes:

* [2018 Munich](https://2018.igem.org/Team%3AMunich/Notebook)
* [2014 ATOMS-Turkiye](https://2014.igem.org/Team%3AATOMS-Turkiye/Notebook)
* [2014 Tec Monterrey](https://2014.igem.org/Team%3ATec-Monterrey/ITESM14_project.html#tab_notebook)
* [2014 Kyoto](https://2014.igem.org/Team%3AKyoto/Notebook/Magnetosome_Formation#title)
* [2014 Cornell](https://2014.igem.org/Team%3ACornell/notebook)

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:OSA/Engineering

![](https://static.igem.org/mediawiki/2020/a/ab/T--OSA--home--logo.png)

* [Home](https://2020.igem.org/Team%3AOSA)
* TEAMS
  + [Team
    Introduction](https://2020.igem.org/Team%3AOSA/Team_Introduction)
  + [Collaboration](https://2020.igem.org/Team%3AOSA/Collaborations)
  + [Attribution](https://2020.igem.org/Team%3AOSA/Attributions)
* PROJECT
  + [Description](https://2020.igem.org/Team%3AOSA/Description)
  + [DNAzyme](https://2020.igem.org/Team%3AOSA/DNAzyme)
  + [Signal
    Amplification System](https://2020.igem.org/Team%3AOSA/Signal_Amplification_System)
  + [DNAzyme
    Mutation](https://2020.igem.org/Team%3AOSA/DNAzyme_Mutation)
  + [Proof
    of Concept](https://2020.igem.org/Team%3AOSA/Proof_Of_Concept)
  + [Model](https://2020.igem.org/Team%3AOSA/Model)
  + [Engineering
    Success](https://2020.igem.org/Team%3AOSA/Engineering)
* Human
  Practice
  + [Integrated
    Human Practice](https://2020.igem.org/Team%3AOSA/Human_Practices)
  + [Public
    Engagement](https://2020.igem.org/Team%3AOSA/Public_Engagement)
  + [Proposed
    Implementation](https://2020.igem.org/Team%3AOSA/Implementation)
* [LAB](https://2020.igem.org/Team%3AOSA/Model)
  + [Safety](https://2020.igem.org/Team%3AOSA/Safetys)
  + [Notebook](https://2020.igem.org/Team%3AOSA/Notebooks)
  + [Protocols](https://2020.igem.org/Team%3AOSA/Protocols)
* [PARTS](https://2020.igem.org/Team%3AOSA/PARTS)
* [COMPETITION](https://2020.igem.org/Team%3AOSA/COMPETITION)
* [JUDGING](https://2020.igem.org/Team%3AOSA/Safety)
  + [Contribution](https://2020.igem.org/Team%3AOSA/Contribution)
  + [Medal](https://2020.igem.org/Team%3AOSA/JUDGING_Medal)

After individual testings of signal amplification systems, we
conducted experiments on TO-DNA and CRISPR-Cas12a.

# Individual Testing:

First, we conducted signal input testing for the two systems. The aim
of this testing is to determine the lowest concentration of key
strand/ssDNA required to trigger reaction.

![](https://static.igem.org/mediawiki/2020/3/34/T--OSA--Engineering_Success_1.png)

**Fig 2.7.1** TO-DNA signal input testing with different concentrations
of key strand 1.

![](https://static.igem.org/mediawiki/2020/4/49/T--OSA--Engineering_Success_2.png)

**Fig 2.7.2** CRISPR-Cas12a signal input testing with different
concentrations of ssDNA-15nt.

Both TO-DNA and CRISPR-Cas12a are successful in their individual
testing. The lower limit of key strand 1 is 1 nM, and the limit of
ssDNA-15nt is 5 nM.

# Simulated Combination Testing:

First, we conducted signal input testing for the two systems. The aim
of this testing is to determine whether the two systems are suitable
to bind to DNAzyme.

![](https://static.igem.org/mediawiki/2020/b/b3/T--OSA--Engineering_Success_3.png)

**Fig 2.7.3** TO-DNA simulated combination testing with complementary
sequences.

![](https://static.igem.org/mediawiki/2020/2/27/T--OSA--Engineering_Success_4.png)

**Fig 2.7.4** CRISPR-Cas12a simulated combination testing with dsDNA.

From TO-DNA testing, we discovered that key strand sequence is too
long to be paired with its complementary sequence. The difference in
length results in early reaction, since the extended part of key
strand will bind to TO-DNA before DNAzyme cleaves its substrate.
Therefore, fluorescence will be emitted despite the absence of heavy
metal ions.

From CRISPR-Cas12a testing, we determined that the Cas12a reaction
will only be triggered by single strand sequence. Thus, we selected
CRISPR-Cas12a as a prospective system for combination with DNAzymes.

![](https://static.igem.org/mediawiki/2020/f/fa/T--OSA--home--footer.png)

[![](https://static.igem.org/mediawiki/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:OSA/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.

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).

### What should this page contain?

* Protocols
* Experiments

### 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/2020/8/8b/HQ_page_logo.jpg)](https://2020.igem.org)

# Team:OSA/Results

# Results

You can describe the results of your project and your future plans here.

### 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.

### 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)