iGEM 2019 — Team UPRM

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# Team:UPRM/Notebook

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[HOME](https://2019.igem.org/Team%3AUPRM)

TEAM

[Team Members](https://2019.igem.org/Team%3AUPRM/Team)
[Collaborations](https://2019.igem.org/Team%3AUPRM/Collaborations)

PROJECT

[Description](https://2019.igem.org/Team%3AUPRM/Description)
[Design](https://2019.igem.org/Team%3AUPRM/Design)
[Experiments & Results](https://2019.igem.org/Team%3AUPRM/Experiments)
[Notebook](https://2019.igem.org/Team%3AUPRM/Notebook)
[Contribution](https://2019.igem.org/Team%3AUPRM/Contribution)
[Attributions](https://2019.igem.org/Team%3AUPRM/Attributions)

[SAFETY](https://2019.igem.org/Team%3AUPRM/Safety)

HUMAN PRACTICES

[Integrated Human Practices](https://2019.igem.org/Team%3AUPRM/Human_Practices)
[Education & Engagement](https://2019.igem.org/Team%3AUPRM/Public_Engagement)

AWARDS

[Medals](https://2019.igem.org/Team%3AUPRM/Medals)

[JUDGING FORM ⇗](https://igem.org/2019_Judging_Form?team=UPRM)

# Notebook

### June

June 25-31

### July

July 1-8July 9-16July 17-24July 25-31

### August

August 1-8August 9-16August 17-24August 25-31

### September

September 1-8September 9-16September 17-24September 25-30

# Protocols

Protocols

##### Thanks to our sponsors

![](https://static.igem.org/mediawiki/2019/3/37/T--UPRM--spon.jpeg)


[![](https://2019.igem.org/wiki/images/8/8b/HQ_page_logo.jpg)](https://2019.igem.org)

# Team:UPRM/Experiments

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[HOME](https://2019.igem.org/Team%3AUPRM)

TEAM

[Team Members](https://2019.igem.org/Team%3AUPRM/Team)
[Collaborations](https://2019.igem.org/Team%3AUPRM/Collaborations)

PROJECT

[Description](https://2019.igem.org/Team%3AUPRM/Description)
[Design](https://2019.igem.org/Team%3AUPRM/Design)
[Experiments & Results](https://2019.igem.org/Team%3AUPRM/Experiments)
[Notebook](https://2019.igem.org/Team%3AUPRM/Notebook)
[Contribution](https://2019.igem.org/Team%3AUPRM/Contribution)
[Attributions](https://2019.igem.org/Team%3AUPRM/Attributions)

[SAFETY](https://2019.igem.org/Team%3AUPRM/Safety)

HUMAN PRACTICES

[Integrated Human Practices](https://2019.igem.org/Team%3AUPRM/Human_Practices)
[Education & Engagement](https://2019.igem.org/Team%3AUPRM/Public_Engagement)

AWARDS

[Medals](https://2019.igem.org/Team%3AUPRM/Medals)

[JUDGING FORM ⇗](https://igem.org/2019_Judging_Form?team=UPRM)

# **Objectives:**

* Perform digestion procedure according to the preset cloning plan.
* Fragments and vectors digestion in order to obtain the specific DNA to the ligation procedure.
* Prototype assembly
* Successful transformation of the prototype in *Escherichia coli.*

We have built a complete Cloning protocol in order to achieve our prototype assembly.

# Wet Lab

#### First Stage

The main objective is to successfully digest the seven fragments necessary to complete the prototype assembly.

![](https://static.igem.org/mediawiki/2019/1/1c/T--UPRM--1.png)

| Fragment | Stock Concentration |
| --- | --- |
| 1a | 200 ng/µL |
| 1b | 200 ng/µL |
| 1c | 200 ng/µL |
| 2 | 200 ng/µL |
| 3 | 200 ng/µL |
| 4 | 200 ng/µL |
| 5 | 200 ng/µL |

The fragments was digested using EcoRI and PstI restriction enzymes. Digestion reactions were calculated for each fragment. After digestion enzymes were inactivated at 20 °C (EcoRI) and 80°C (PstI). Then, an electrophoresis gel was prepared to analysed the digested fragments.

# Protocol specifications:

### Digestion Reaction

1. Mix in a 1.5 mL microcetrifuge tube

* 5μl DNA (土200 ng/ μl)
* 0.5 μl PstI
* 0.5 μl EcoRI
* 1 μl enzyme buffer
* 3 μl ddH2O

10 μl Total Volume

2. Incubate at 37 °C for 15 min to 2 hrs.
3. Inactivate enzyme at 60 °C for 20 min and 80 °C for 20 mins.

Our ideal working concentration is over 100 ng/uL.

![](https://static.igem.org/mediawiki/2019/7/7c/T--UPRM--Results.png)

**Figure 2**: Fragments digestion. In the electrophoresis gel (agarose 1%) strong fragment concentrations are notable. To see a fragment in a gel a minimum of a 100 ng is needed. The observed results indicate that a concentration of 土150 ng were conserved along the process due to the bands brightness.

## SynBio 101 Results

[#### Survey Results](https://2019.igem.org/Team%3AUPRM/Surveys)

Surveys were used as the instrument to gather quantitative information that represents the general public’s knowledge about Synthetic Biology, Genetically Modified Organisms (GMOs) and agriculture. This helped us have a better understanding of how obstacles in these topics could be targeted using our prototype. After interviewing 53 people, in summary, their answers regarding their knowledge about Synthetic Biology, their consumption of GMOs, and if they think the field is being addressed adequately by the general public:

![](https://static.igem.org/mediawiki/2019/8/88/T--UPRM--pie1.png)

![](https://static.igem.org/mediawiki/2019/9/99/T--UPRM--pie2.png)

![](https://static.igem.org/mediawiki/2019/e/e6/T--UPRM--pie3.png)

[#### Workshop Results](https://2019.igem.org/Team%3AUPRM/Operation456)

![](https://static.igem.org/mediawiki/2019/a/ae/T--UPRM--prepostgraph.png)

During the Operation 456 Workshops we wanted to track the improvement in knowledge of the 35 participants. For this, students were given pre and post tests. We compared the results of these tests and saw an increase of correct answers from 41.43% to 78.93%, showing an improvement of 37.5%.

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# Team:UPRM/Results

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[HOME](https://2019.igem.org/Team%3AUPRM)

TEAM

[Team Members](https://2019.igem.org/Team%3AUPRM/Team)
[Collaborations](https://2019.igem.org/Team%3AUPRM/Collaborations)

PROJECT

[Description](https://2019.igem.org/Team%3AUPRM/Description)
[Design](https://2019.igem.org/Team%3AUPRM/Design)
[Experiments & Results](https://2019.igem.org/Team%3AUPRM/Experiments)
[Notebook](https://2019.igem.org/Team%3AUPRM/Notebook)
[Contribution](https://2019.igem.org/Team%3AUPRM/Contribution)
[Attributions](https://2019.igem.org/Team%3AUPRM/Attributions)

[SAFETY](https://2019.igem.org/Team%3AUPRM/Safety)

HUMAN PRACTICES

[Integrated Human Practices](https://2019.igem.org/Team%3AUPRM/Human_Practices)
[Education & Engagement](https://2019.igem.org/Team%3AUPRM/Public_Engagement)

AWARDS

[Medals](https://2019.igem.org/Team%3AUPRM/Medals)

[JUDGING FORM ⇗](https://igem.org/2019_Judging_Form?team=UPRM)

![]()

# 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 also be on the part's Main Page on the Registry.** 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)