iGEM 2019 — Team UA_Huntsville

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# Team:UA Huntsville/Notebook

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

TEAM

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

PROJECT

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

PARTS

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

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

HUMAN PRACTICES

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

AWARDS

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

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

# Notebook

## March

Held interest meetings and recruited members

Registered as an iGEM team

## April

Administrative work

## May

Began weekly summer planning meetings

Brainstorming began on projects

Trained members in lab safety

## June

Selected Cosmicoli as our project

Made first contact with our iGEM mentor

## July

Hosted a professional communication workshop

hosted a fun Spirited Art Fundraiser

Received IR9-50-1 and Founder Delta\_e14 from Cox Lab, reproduced literature results.

Designed Expression Systems for Dsup and CAHS

## August

Registered for the Jamboree!

Initial experiments for transforming parts into vector chassis, MC1061.

Digestions and Test Ligations

## September

Digestions and test Ligations

Started Ligations! Encountered troubles.

## October

Ligations started using Red White Screening, led to isolation of likely pBAD/AraC-Dsup and pTet-CAHS.

Isolation and Dsup and CAHS expression systems.


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# Team:UA Huntsville/Experiments

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

TEAM

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

PROJECT

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

PARTS

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

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

HUMAN PRACTICES

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

AWARDS

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

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

## Research

## Protocols

Workshop 1 Handout
Basic protocols

[Basic Protocols](https://static.igem.org/mediawiki/2019/1/1f/T--UA_Huntsville--Workshop_1_handout.pdf)

Pouring Plate and Culturing Protocols:

[Pouring Plate and Culturing Protocols](https://static.igem.org/mediawiki/2019/c/c6/T--UA_Huntsville--Plate_Preparation_and_Culturing_Protocols.pdf)

Transformation Protocols

[Transformation Protocols](https://static.igem.org/mediawiki/2019/6/65/T--UA_Huntsville--Transformation_Protocols.pdf)

Alkaline Lysis Mini Prep Protocols:

[Alkaline Lysis Mini Prep Protocols](https://static.igem.org/mediawiki/2019/8/8e/T--UA_Huntsville--Alkaline_Lysis_Mini_Prep_Protocol.pdf)

PCR Protocol
PCR protocols were conducted as listed on NEB website:

[PCR Protocol](https://www.neb.com/protocols/0001/01/01/taq-dna-polymerase-with-standard-taq-buffer-m0273)

Litigation Protocols
Litigation followed protocols as listed by New England Biolabs. Instead of masses listed, 0.02pmol vector and 0.06 pmol insert were used to fine-tune to each vector and insert. Protocol may be found at the following link:

[Litigation Protocols](https://www.neb.com/protocols/1/01/01/dna-ligation-with-t4-dna-ligase-m0202)

## Experiments

Experiment description can be found in the following pdf:

[Experiments](https://static.igem.org/mediawiki/2019/c/c4/T--UA_Huntsville--experimental_design.pdf)

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# Team:UA Huntsville/Results

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

TEAM

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

PROJECT

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

PARTS

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

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

HUMAN PRACTICES

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

AWARDS

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

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

# Project Results

#### Results Thus Far

pBAD/AraC-Dsup and pTet-CAHS constructs have been successfully synthesized. These constructs contain the key tardigrade radiotolerance genes that will become key components of the final plasmid. Digestion of 2xTer-Amp backbone will be carried out shortly, and then the two constructs will be ligated to this backbone. Once this has been achieved, the two plasmid constructs will be transformed into a LacZ frameshift, exposed to Ultraviolet radiation, and the amount of revertants vs. empty vector will be assessed. These experiments are in process at the time of completion of the website.

#### Future Directions

In the future, we intend to test these constructs transformed into IR9-50-1 strain and measure survival under UV radiation and Ionizing radiation. We also intend to test the effects of microgravity on the viability and growth rates of these cells using a microgravity bioreactor from Synthecon. It is our hope that the cosmicoli will maintain radiotolerance and fitness in microgravity to confirm their effectiveness for space-based research.