iGEM 2019 — Team UAAAN

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# Team:UAAAN/Notebook

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* Team
  + [Team Members](https://2019.igem.org/Team%3AUAAAN/Team)
  + [Collaborations](http://2019.igem.org/Team%3AUAAAN/Collaborations)
  + [Attributions](https://2019.igem.org/Team%3AUAAAN/Attributions)
* Project
  + [Description](http://2019.igem.org/Team%3AUAAAN/Description)
  + [Design](http://2019.igem.org/Team%3AUAAAN/Design)
  + [Notebook](http://2019.igem.org/Team%3AUAAAN/Notebook)
  + [InterLab](http://2019.igem.org/Team%3AUAAAN/InterLab)
  + [Safety](http://2019.igem.org/Team%3AUAAAN/Safety)
  + [Demonstrate](http://2019.igem.org/Team%3AUAAAN/Demonstrate)
* Parts
  + [Parts](http://2019.igem.org/Team%3AUAAAN/Parts)
  + [Basic Parts](http://2019.igem.org/Team%3AUAAAN/Basic_Part)
  + [Composite Parts](http://2019.igem.org/Team%3AUAAAN/Composite_Part)
  + [Part Collection](http://2019.igem.org/Team%3AUAAAN/Part_Collection)
* Human Practices
  + [Human Practices](https://2019.igem.org/Team%3AUAAAN/Human_Practices)
  + [Public Engagement](https://2019.igem.org/Team%3AUAAAN/Public_Engagement)
* Awards
  + [Medal Checklist](http://2019.igem.org/Team%3AUAAAN/Awards)

[Previous](#ua-background)

# Welcome to team: **UAAAN**

Universidad Autónoma Agraria Antonio Narro

[Next](#ua-background)

## Abstract

## Project

[Description](http://2019.igem.org/Team%3AUAAAN/Description)

[Notebook](https://2019.igem.org/Team%3AUAAAN/Notebook)

[INFO LANGUAGES](https://www.youtube.com/watch?v=ghDXxD5_w1E&t=2s)

## Fun GALERY

![](https://2019.igem.org/wiki/images/7/72/T--UAAAN--equipo.jpg)

![](https://2019.igem.org/wiki/images/3/3e/T--UAAAN--Blopers_iGEM_UAAAN.jpg)

![](https://2019.igem.org/wiki/images/b/b5/T--UAAAN--Blopers_iGEM_UAAAN2.jpg)

![](https://2019.igem.org/wiki/images/b/b3/T--UAAAN--Bloopers_iGEM_UAAAN.jpg)

![](https://2019.igem.org/wiki/images/f/f6/T--UAAAN--Bloopers_iGEM_UAAAN3.jpg)

![](https://2019.igem.org/wiki/images/7/7a/T--UAAAN--Bloopers_iGEM_UAAAN4.jpg)

The laughter never failed!!!

![](https://2019.igem.org/wiki/images/a/a5/T--UAAAN--Bloopers_iGEM_UAAAN5.jpg)
![](https://2019.igem.org/wiki/images/0/08/T--UAAAN--Bloopers_iGEM_UAAAN6.jpg)
![](https://2019.igem.org/wiki/images/d/d8/T--UAAAN--Bloopers_iGEM_UAAAN7.jpg)
![](https://2019.igem.org/wiki/images/1/13/T--UAAAN--Bloopers_iGEM_UAAAN11.jpg)![](https://2019.igem.org/wiki/images/d/de/T--UAAAN--Bloopers_iGEM_UAAAN16.jpg)
![](https://2019.igem.org/wiki/images/f/f7/T--UAAAN--Bloopers_iGEM_UAAAN25.jpg)
![](https://2019.igem.org/wiki/images/f/f2/T--UAAAN--Bloopers_iGEM_UAAAN26.jpg)
![](https://2019.igem.org/wiki/images/7/75/T--UAAAN--Bloopers_iGEM_UAAAN30.jpg)
![](https://2019.igem.org/wiki/images/a/ae/T--UAAAN--Bloopers_iGEM_UAAAN_34.jpg)
![]()
![](https://2019.igem.org/wiki/images/5/53/T--UAAAN--Bloopers_iGEM_UAAAN_38.jpg)
![](https://2019.igem.org/wiki/images/d/d7/T--UAAAN--Bloopers_iGEM_UAAAN_39.jpg)

## Special thanks to all our sponsors!

![](http://2017.igem.org/wiki/images/7/7d/T--UAlberta--ThermoFisher_Logo.png)

![](http://2017.igem.org/wiki/images/6/62/T--UAlberta--IDT_Logo.png)

![](http://2017.igem.org/wiki/images/b/b7/T--UAlberta--SnapGene_Logo.png)

![](http://2017.igem.org/wiki/images/5/54/T--UAlberta--UofA_Logo.png)

![](http://2017.igem.org/wiki/images/5/59/T--UAlberta--geekStarter_Logo.png)

![](http://2017.igem.org/wiki/images/d/d2/T--UAlberta--Qiagen_Logo.png)

![](http://2017.igem.org/wiki/images/d/d0/T--UAlberta--NEB_Logo.png)

![](http://2017.igem.org/wiki/images/0/0d/T--UAlberta--Agilent_Logo.png)

![](http://2017.igem.org/wiki/images/0/00/T--UAlberta--URI_Logo.png)

![](http://2017.igem.org/wiki/images/3/38/T--UAlberta--Mathworks_Logo.png)

![](http://2017.igem.org/wiki/images/3/3d/T--UAlberta--Joeys_Logo.png)

## Social Media

###### igem.ualberta@gmail.com


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

# Team:UAAAN/Experiments

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HOME

TEAM

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

PROJECT

[Description](https://2019.igem.org/Team%3AUAAAN/Description)
[Design](https://2019.igem.org/Team%3AUAAAN/Design)
[Experiments](https://2019.igem.org/Team%3AUAAAN/Experiments)
[Notebook](https://2019.igem.org/Team%3AUAAAN/Notebook)
[Contribution](https://2019.igem.org/Team%3AUAAAN/Contribution)
[Results](https://2019.igem.org/Team%3AUAAAN/Results)
[Demonstrate](https://2019.igem.org/Team%3AUAAAN/Demonstrate)
[Improve](https://2019.igem.org/Team%3AUAAAN/Improve)

PARTS

[Parts Overview](https://2019.igem.org/Team%3AUAAAN/Parts)
[Basic Parts](https://2019.igem.org/Team%3AUAAAN/Basic_Part)
[Composite Parts](https://2019.igem.org/Team%3AUAAAN/Composite_Part)
[Part Collection](https://2019.igem.org/Team%3AUAAAN/Part_Collection)

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

HUMAN PRACTICES

[Human Practices](https://2019.igem.org/Team%3AUAAAN/Human_Practices)
[Education & Engagement](https://2019.igem.org/Team%3AUAAAN/Public_Engagement)
[Integrative Human Practices](https://2019.igem.org/Team%3AUAAAN/Integrative%20Human%20Practices)

AWARDS

[Entrepreneurship](https://2019.igem.org/Team%3AUAAAN/Entrepreneurship)
[Hardware](https://2019.igem.org/Team%3AUAAAN/Hardware)
[Measurement](https://2019.igem.org/Team%3AUAAAN/Measurement)
[Model](https://2019.igem.org/Team%3AUAAAN/Model)
[Plant](https://2019.igem.org/Team%3AUAAAN/Plant)
[Software](https://2019.igem.org/Team%3AUAAAN/Software)

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

Document

* [Protocols: Practice Planning](#seccion1)
* [Basic laboratory techniques](#seccion2)
* [Incubation](#seccion3)
* [Thermal shock](#seccion4)
* [Seeding of transformed cells and control cells](#seccion5)
* [Thermal shock](#seccion6)

### Practice Planning

Each of the three hands-on sessions is scheduled to be held at 50-minute intervals in succession.

## Proposed program for the transformation of E. coli (DH5α)

Day 1: Introduction Reading and discussion of considerations
Day 2: Transformation: Transform cells and sow plates.
Day 3: Collect and analyze results.
Observed transformed cells and controls.
Day 4: Analyze and interpret results.
Discussion and complementary activities.

# Sterilization technique

In any microbiological technique it is essential not to introduce contaminating bacteria into the experiment. Since the contaminating bacteria are found everywhere (fingers, table, etc.) it is important to prevent the material from coming into contact with these surfaces. When students are working they should avoid touching the end of the sowing handle, the tip of the pipette and the agar of the plate and should also avoid contact with any contaminated surface. For various reasons, very small quantities and volumes are used in the laboratory, requiring the use of specialized equipment such as analytical scales and micropipettes.

# Working with E. coli

The organism used in this process, a strain of E. coli (DH5α), the vector that contains the gene of our interest, and the transformed cells that are formed by combination of the above, are not pathogenic organisms such as E. coli O157:H7, which is known for its involvement in food-poisoning infections. However, the handling of E. coli (DH5α) must be done according to standard microbiological safety standards. Among these we can cite:
• Work surfaces are disinfected once a day and each time an accidental spill occurs (with ethanol). • All liquid or solid waste must be disinfected before it is thrown away.
• Hands should be washed: or after handling material with recombinant DNA molecules, and or before leaving the laboratory.
• All procedures should be performed with caution to minimize aerosol formation.
• Micropipettes shall be used, it is forbidden to pipette with the mouth.
• It is forbidden to eat, drink, smoke and apply cosmetics in the work area.
• Protective glasses and gloves are recommended.

# Disinfection and removal of material

With the help of the autoclave, sterilize all used material, then double bag the single-use and discard. Wash well those that are not disposable. Do not throw the agar into the pipe.

This protocol is designed to use an incubator at 37 ºC. The transformation can be done without using the incubator, but then the number of days needed for the colonies to grow will depend on the ambient temperature. The best results are observed with crops of 24-48 hours and with colonies with measures of 1-1.5 mm of diameter. Cooling the plates will significantly reduce the processing efficiency. The optimal growth temperature of E. coli is 37 ºC and lower temperatures will produce a lower growth rate. At 28 ºC, 2 days of incubation are needed to obtain colonies of appropriate size. At 21 ºC, 3 days of incubation are required.

# Methodological indications

Transfer of bacterial colonies from the agar plates to the tubes When you take a colony from the culture plates, you may be tempted to take more cells than necessary. It should be remembered that a single colony with a diameter of 1 mm contains millions of cells.

The procedure used to promote foreign DNA collection is called thermal shock. It is important to follow the recommended times. Also important is the sudden change in temperature and the duration of the thermal shock. For optimal results, the tubes with the cell suspension should be removed directly from the ice, placed in a bath at 42 ºC for 50 seconds and immediately put on ice again. For example, if there is no thermal shock, one tenth of transformations will occur, or if the duration of the shock is 90 seconds, half of transformations will be obtained than with 50 seconds.

Adding too much transformation medium to plates is counterproductive as plates cannot absorb excess fluid and seeding will be uneven. The passage of bacterial suspension from tubes to petri dishes should be done with care. The bacteria are deposited at the bottom of the tubes, so the tubes should be shaken. To do this you must take the top of the tube with your thumb and index fingers and you must tap the bottom of the tube with the index finger of the other hand. You should also make sure that the petri dishes are taped once the bacterial suspension has been extended.

# Transformation solution

The Ca 2+ ions present in the solution (Cacl2) neutralise the repulsion between the negative charges of the DNA phosphate groups and the cell membrane phospholipids, allowing DNA to enter the cells.

Thermal shock increases cell membrane permeability to DNA. Although it is not known why, the duration of the thermal shock is known to be critical to the process and has been optimised for the type of bacteria and conditions used.

# Recovery

The 10 minutes of incubation after the addition of the LB broth allows the cells to grow and express the antibiotic resistance gene, and so the transformed cells survive in the antibiotic plates, which allows to know if the vector has been integrated and is expressing itself. Incubation can be carried out at room temperature or in the incubator at 37 ºC to increase processing efficiency by up to 10 times.

</html>

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# Team:UAAAN/Results

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HOME

TEAM

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

PROJECT

[Description](https://2019.igem.org/Team%3AUAAAN/Description)
[Design](https://2019.igem.org/Team%3AUAAAN/Design)
[Experiments](https://2019.igem.org/Team%3AUAAAN/Experiments)
[Notebook](https://2019.igem.org/Team%3AUAAAN/Notebook)
[Contribution](https://2019.igem.org/Team%3AUAAAN/Contribution)
[Results](https://2019.igem.org/Team%3AUAAAN/Results)
[Demonstrate](https://2019.igem.org/Team%3AUAAAN/Demonstrate)
[Improve](https://2019.igem.org/Team%3AUAAAN/Improve)

PARTS

[Parts Overview](https://2019.igem.org/Team%3AUAAAN/Parts)
[Basic Parts](https://2019.igem.org/Team%3AUAAAN/Basic_Part)
[Composite Parts](https://2019.igem.org/Team%3AUAAAN/Composite_Part)
[Part Collection](https://2019.igem.org/Team%3AUAAAN/Part_Collection)

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

HUMAN PRACTICES

[Human Practices](https://2019.igem.org/Team%3AUAAAN/Human_Practices)
[Education & Engagement](https://2019.igem.org/Team%3AUAAAN/Public_Engagement)
[Integrative Human Practices](https://2019.igem.org/Team%3AUAAAN/Integrative%20Human%20Practices)

AWARDS

[Entrepreneurship](https://2019.igem.org/Team%3AUAAAN/Entrepreneurship)
[Hardware](https://2019.igem.org/Team%3AUAAAN/Hardware)
[Measurement](https://2019.igem.org/Team%3AUAAAN/Measurement)
[Model](https://2019.igem.org/Team%3AUAAAN/Model)
[Plant](https://2019.igem.org/Team%3AUAAAN/Plant)
[Software](https://2019.igem.org/Team%3AUAAAN/Software)

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

# 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)