iGEM 2018 — Team East_Chapel_Hill

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# Team:East Chapel Hill/Notebook

[East Chapel Hill](https://2018.igem.org/Team%3AEast_Chapel_Hill)

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  + [Description](https://2018.igem.org/Team%3AEast_Chapel_Hill/Description)
  + [Results](https://2018.igem.org/Team%3AEast_Chapel_Hill/Results)
  + [Measurement](https://2018.igem.org/Team%3AEast_Chapel_Hill/Measurement)
* [Lab](#!)
  + [Experiments](https://2018.igem.org/Team%3AEast_Chapel_Hill/Experiments)
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* [Human Practices](#!)
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  + [Public Engagement](https://2018.igem.org/Team%3AEast_Chapel_Hill/Public_Engagement)

# Lab Notebook 2018

#### [**June 26th, 2018 through June 29th, 2018**](#collapse1)

#### [**July 5th, 2018 through July 13th, 2018**](#collapse4)

#### [**August 1st, 2018 through August 10th, 2018**](#collapse5)

#### [**August 22nd, 2018**](#collapse3)


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# Team:East Chapel Hill/Experiments

[East Chapel Hill](https://2018.igem.org/Team%3AEast_Chapel_Hill)

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  + [Description](https://2018.igem.org/Team%3AEast_Chapel_Hill/Description)
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  + [Measurement](https://2018.igem.org/Team%3AEast_Chapel_Hill/Measurement)
* [Lab](#!)
  + [Experiments](https://2018.igem.org/Team%3AEast_Chapel_Hill/Experiments)
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# Experiments

#### [Gel Extraction](#P1-collapse)

![](https://static.igem.org/mediawiki/2018/b/bf/T--East_Chapel_Hill--GELEXTRACT.png)

#### [Chemically Competent Cells](#P2-collapse)

![](https://static.igem.org/mediawiki/2018/a/a1/T--East_Chapel_Hill--CHEMICALLYCOMPETENT.png)

#### [Serial Dilutions](#P3-collapse)

![](https://static.igem.org/mediawiki/2018/4/42/T--East_Chapel_Hill--SERIALDILUTIONS.png)

#### [Gibson Cloning and Heat Shock](#P4-collapse)

![](https://static.igem.org/mediawiki/2018/a/ad/T--East_Chapel_Hill--GIBSONHEATSHOCK-2_%28dragged%29.png)

#### [Agar Gel Electrophoresis](#P5-collapse)

![](https://static.igem.org/mediawiki/2018/c/c7/T--East_Chapel_Hill--AGARGELELECTRO.png)

#### [Polymerase Chain Reaction](#P6-collapse)

![](https://static.igem.org/mediawiki/2018/0/07/T--East_Chapel_Hill--PCR.png)

#### [Glass Bead Plating](#P7-collapse)

![](https://static.igem.org/mediawiki/2018/2/2f/T--East_Chapel_Hill--GLASSBEAD.png)

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# Team:East Chapel Hill/Results

[East Chapel Hill](https://2018.igem.org/Team%3AEast_Chapel_Hill)

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  + [Measurement](https://2018.igem.org/Team%3AEast_Chapel_Hill/Measurement)
* [Lab](#!)
  + [Experiments](https://2018.igem.org/Team%3AEast_Chapel_Hill/Experiments)
  + [Safety](https://2018.igem.org/Team%3AEast_Chapel_Hill/Safety)
  + [Notebook](https://2018.igem.org/Team%3AEast_Chapel_Hill/Notebook)
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# Results

 Due to experimental errors and lack of reproducibility, there is some uncertainty in several of our plating assays. However, the results from August 22nd, 2018 appear to be the most accurate due to the lack of growth of the fluoride-binding mutant. Upon looking at these results, we can reasonably infer that the promoter J23\_102 and the riboswitch variation ‘FRS1’ are most effective in allowing for dynamic bacterial growth across varied levels of fluoride.

![](https://static.igem.org/mediawiki/2018/9/98/T--East_Chapel_Hill--RESULTSPAGE1.png)
![](https://static.igem.org/mediawiki/2018/0/08/T--East_Chapel_Hill--RESULTSPAGE2.png)

# Future Directions

Short Term:

 - We hope to repeat our plating assays to gain more conclusive results in terms of ensuring a direct correlation between the fluoride concentration and bacterial growth

 - The CHOP system can be used to screen and select fluoride riboswitches with a higher affinity for fluoride, leading to more growth at lower levels of fluoride and more effective detection. This would also make the system more valuable for implementation in communities where a fluoride detection system is most needed.

 - We hope to test the effectiveness of a liquid culture for our construct. Different riboswitch promoter pairs can be inserted into CHOP and response to varying levels of fluoride can be measured.

Long Term:

 - In order to bioremediate excess levels of fluoride, we envision using fluoride riboswitches to express genes that will allow for enzymes to metabolize fluoride in the body.

 - To sequester fluoride we hope to identify fluoride riboswitches with a higher affinity to fluoride and attach multiple copies of the riboswitches to nanoparticles

 - For easier implementation, we hope to allow riboswitches to regulate a reporter for a more obvious sign when unhealthy levels of fluoride are present in drinking water.

 - CHOP can be used to evaluate other transcriptional riboswitches that bind different metal ions, such as cobalt or nickel.