iGEM 2019 — Team Lubbock_TTU

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

# Team:Lubbock TTU/Notebook

[![](https://static.igem.org/mediawiki/2019/c/c0/T--Lubbock_TTU--Logonoyear_2019.png)](https://2019.igem.org/Team%3ALubbock_TTU)

Judging

[Judging Form](https://igem.org/2019_Judging_Form?id=2919)

[Medal Requirements](https://2019.igem.org/Team%3ALubbock_TTU/Medal)

Sponsors

[Sponsors](https://2019.igem.org/Team%3ALubbock_TTU/Sponsors)

Team

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

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

[Attributions](https://2019.igem.org/Team%3ALubbock_TTU/Attributions)

Application

[Product Design](https://2019.igem.org/Team%3ALubbock_TTU/Product_Design)

[Modeling](https://2019.igem.org/Team%3ALubbock_TTU/Model)

Human Practices

[Human Practices](https://2019.igem.org/Team%3ALubbock_TTU/Human_Practices)

[Integrated Human Practices](https://2019.igem.org/Team%3ALubbock_TTU/Integrated_Human_Practices)

Lab

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

[Protocols](https://2019.igem.org/Team%3ALubbock_TTU/Protocols)

[Results](https://2019.igem.org/Team%3ALubbock_TTU/Results)

[Demonstrate](https://2019.igem.org/Team%3ALubbock_TTU/Demonstrate)

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

Project

[Description](https://2019.igem.org/Team%3ALubbock_TTU/Description)

[inspiration](https://2019.igem.org/Team%3ALubbock_TTU/inspiration)

[Design](https://2019.igem.org/Team%3ALubbock_TTU/Design)

[Parts](https://2019.igem.org/Team%3ALubbock_TTU/Parts)

## Lab Notebook

![](https://static.igem.org/mediawiki/2019/9/95/T--Lubbock_TTU--Notebookred_2019.png)

####

##### JANUARY - MAY

Team meetings and preparation for wet lab work.

 Read papers:

o Tropane and Granatane Alkaloid Biosynthesis: A Systematic Analysis

o Metabolic Engineering of Escherichia coli for the Production of Putrescine: A Four Carbon Diamine

o Tropinone synthesis via an atypical polyketide synthase and P450-mediated cyclization

o Loop assembly: a simple and open system for recursive fabrication of DNA circuits

o CIDAR MoClo: Improved MoClo Assembly Standard and New E. coli Part Library Enable Rapid Combinatorial Design for Synthetic and Traditional Biology

o Systems Metabolic Engineering Strategies: Integrating Systems and Synthetic Biology with Metabolic Engineering

o Scale-up of industrial microbial processes

o Synthetic and systems biology for microbial production of commodity chemicals

o Engineering Cellular Metabolism

o Automated Parameterization of Predictive Kinetic Metabolic Models from Sparse Datasets for Efficient Optimization of
Many-Enzyme Heterologous Pathways

##### APRIL

Team presented at Texas Tech University Undergraduate Research Conference.

##### MAY

Team received BL21-OP-A3 E.coli strain, which produces N-methylpyrrolinum, from Shanghai Institute of Plant Physiology.

##### JUNE 8th

ULabs Workshop.

##### JUNE 22nd

ULabs Workshop.

##### JULY

Plated and confirmed putrescine producing platform strain, XQ52..

##### JULY 7th

pOdd-1 juniperGFP transcriptional unit construction and confirmation (successful usage of Loop Assembly method).

##### JULY 27th

Texas iGEM Meetup.

##### JULY 27th

Texas iGEM Meetup.

##### JULY 27th

Texas iGEM Meetup.

##### AUGUST 4th

The pACYCDuet-1 plasmid containing EcODC and AtPMT enzymes and the pET30a plasmids containing AaDAO3 enzymes were mini prep and isolated from the BL21-OP-A3 E.coli strain.

##### AUGUST 5th – 9th

Completed colony PCR on XQ52 strain (not competent cells), and conducted OD 600 measurements using a 250 mL culture shake flask for W3110 (WT of XQ52 strain) competent cell biological test and OD600 E. Coli non-competent XQ52 strain biological test.

##### AUGUST 7th

Sent our commonly used parts to UT for a collaboration.

##### AUGUST 12th – 16th

Completed XQ52 competent cell without Plasmid OD600 for 24+ Hours and W3110 competent cell without plasmid OD600 in a 250 mL shake flask culture.

##### AUGUST 20th – 23rd

Completed OD600 on 96-well-plate for W3110 competent cell and XQ52 competent cell without supplement.

##### AUGUST 21st

Tested UT’s calibration strains using a plate reader.

##### AUGUST 26th – 30th

Completed OD600 on 96-well-plate for W3110 competent cell and XQ52 competent cell without and with supplement.

##### AUGUST 30th

Meeting with UT to discuss collaboration.

##### SEPTEMBER 4th

Transformed pET30a-AaDAO3 into XQ52 competent cells, and utilized PFU PCR to confirm the fragment size of the AaDAO3 gene in the pET30a-AaDAO3 plasmid.

##### SEPTEMBER 11th

Calibration of OD600 and data analysis utilizing microsphere beads with a serial dilution in a 96 well plate.

##### SEPTEMBER 26th

Performed assay to test XQ52 tolerance to tropinone at varying concentrations.

##### SEPTEMBER 29th

Successfully introduced the pACYCDuet-1 plasmid into the XQ52 platform strain containing pET30a plasmid.

##### OCTOBER 3rd

Conversion of pOdd1-2 Loop Assembly plasmids into low copy plasmids which utilize the backbone of the pSB4K5 plasmid.

##### OCTOBER 6th

Successfully introduced the pet-Strep plasmid containing the PYKS enzyme into XQ52 platform straining containing the pACYCDuet-1 and pET30a plasmid (formation of the XQ52-DPS strain).

##### OCTOBER 9th

The XQ52-DPS was induced for grown to an OD of .6 and induced with IPTG and copper sulfate for 24 hours. Oxobutanic acid and oxoglutaric acid were successfully measured using LC-MS.

##### OCTOBER 10th

Team presented at the Center for the Integration of STEM Education and Research (CISER) Research Carnival at Texas Tech University.

##### OCTOBER 11th

Conversion of pOdd3-4 Loop Assembly plasmids into low copy plasmids which utilize the backbone of the pSB4K5 plasmid.


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

# Team:Lubbock TTU/Experiments

[![](https://static.igem.org/mediawiki/2019/c/c0/T--Lubbock_TTU--Logonoyear_2019.png)](https://2019.igem.org/Team%3ALubbock_TTU)

Judging

[Judging Form](https://igem.org/2019_Judging_Form?id=2919)

[Medal Requirements](https://2019.igem.org/Team%3ALubbock_TTU/Medal)

Sponsors

[Sponsors](https://2019.igem.org/Team%3ALubbock_TTU/Sponsors)

Team

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

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

[Attributions](https://2019.igem.org/Team%3ALubbock_TTU/Attributions)

Application

[Product Design](https://2019.igem.org/Team%3ALubbock_TTU/Product_Design)

[Modeling](https://2019.igem.org/Team%3ALubbock_TTU/Model)

Human Practices

[Human Practices](https://2019.igem.org/Team%3ALubbock_TTU/Human_Practices)

[Integrated Human Practices](https://2019.igem.org/Team%3ALubbock_TTU/Integrated_Human_Practices)

Lab

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

[Protocols](https://2019.igem.org/Team%3ALubbock_TTU/Protocols)

[Results](https://2019.igem.org/Team%3ALubbock_TTU/Results)

[Demonstrate](https://2019.igem.org/Team%3ALubbock_TTU/Demonstrate)

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

Project

[Description](https://2019.igem.org/Team%3ALubbock_TTU/Description)

[inspiration](https://2019.igem.org/Team%3ALubbock_TTU/inspiration)

[Design](https://2019.igem.org/Team%3ALubbock_TTU/Design)

[Parts](https://2019.igem.org/Team%3ALubbock_TTU/Parts)

# 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.

Please remember to put all characterization and measurement data for your parts on the corresponding Registry part pages.

### What should this page contain?

* Protocols
* Experiments
* Documentation of the development of your project

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

# Team:Lubbock TTU/Results

[![](https://static.igem.org/mediawiki/2019/c/c0/T--Lubbock_TTU--Logonoyear_2019.png)](https://2019.igem.org/Team%3ALubbock_TTU)

Judging

[Judging Form](https://igem.org/2019_Judging_Form?id=2919)

[Medal Requirements](https://2019.igem.org/Team%3ALubbock_TTU/Medal)

Sponsors

[Sponsors](https://2019.igem.org/Team%3ALubbock_TTU/Sponsors)

Team

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

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

[Attributions](https://2019.igem.org/Team%3ALubbock_TTU/Attributions)

Application

[Product Design](https://2019.igem.org/Team%3ALubbock_TTU/Product_Design)

[Modeling](https://2019.igem.org/Team%3ALubbock_TTU/Model)

Human Practices

[Human Practices](https://2019.igem.org/Team%3ALubbock_TTU/Human_Practices)

[Integrated Human Practices](https://2019.igem.org/Team%3ALubbock_TTU/Integrated_Human_Practices)

Lab

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

[Protocols](https://2019.igem.org/Team%3ALubbock_TTU/Protocols)

[Results](https://2019.igem.org/Team%3ALubbock_TTU/Results)

[Demonstrate](https://2019.igem.org/Team%3ALubbock_TTU/Demonstrate)

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

Project

[Description](https://2019.igem.org/Team%3ALubbock_TTU/Description)

[inspiration](https://2019.igem.org/Team%3ALubbock_TTU/inspiration)

[Design](https://2019.igem.org/Team%3ALubbock_TTU/Design)

[Parts](https://2019.igem.org/Team%3ALubbock_TTU/Parts)

## Results

![](https://static.igem.org/mediawiki/2019/5/5d/T--Lubbock_TTU--Resultsred_2019.png)

* [![Instagram](https://static.igem.org/mediawiki/2019/e/eb/T--MichiganState--InstagramIcon.png)](https://www.instagram.com/ttu_igemraiders/)
* [![Twitter](https://static.igem.org/mediawiki/2019/a/ae/T--MichiganState--TwitterIcon.png)](https://twitter.com/ttuigem?lang=en)
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