Team:Gaston Day School/cadmiumdetector

From 2014.igem.org

(Difference between revisions)
(Created page with "{{CSS/Main}} <html> <head> <link href='http://fonts.googleapis.com/css?family=Open+Sans' rel='stylesheet' type='text/css'> <link href='http://fonts.googleapis.com/css?family...")
 
(11 intermediate revisions not shown)
Line 1: Line 1:
{{CSS/Main}}
{{CSS/Main}}
<html>
<html>
 +
<script>
 +
(function(i,s,o,g,r,a,m){i['GoogleAnalyticsObject']=r;i[r]=i[r]||function(){
 +
(i[r].q=i[r].q||[]).push(arguments)},i[r].l=1*new Date();a=s.createElement(o),
 +
m=s.getElementsByTagName(o)[0];a.async=1;a.src=g;m.parentNode.insertBefore(a,m)
 +
})(window,document,'script','//www.google-analytics.com/analytics.js','ga');
 +
                document.body.className = 'transform'; ga('create', 'UA-53208337-1', 'auto');
 +
ga('send', 'pageview');
 +
</script>
<head>
<head>
<link href='http://fonts.googleapis.com/css?family=Open+Sans' rel='stylesheet' type='text/css'>
<link href='http://fonts.googleapis.com/css?family=Open+Sans' rel='stylesheet' type='text/css'>
Line 15: Line 23:
}
}
-
+
 
-
            [class*='col-'] {
+
[class*='col-'] {
float: left;
float: left;
padding-right: 20px;
padding-right: 20px;
Line 236: Line 244:
#navbar > div > a:active > p.show {
#navbar > div > a:active > p.show {
box-shadow: 0px 0px 25px #888888;
box-shadow: 0px 0px 25px #888888;
-
}
 
-
 
-
.image-wrapper
 
-
{
 
-
position: relative;
 
}
}
-
.scale-image
 
-
{
 
-
height: 100%;
 
-
width: 100%;
 
-
}
 
</style>
</style>
</head>
</head>
-
+
 
-
<body class="transform">
+
<body>
-
+
<div class="row">
<div class="row">
<H1 id="mainhead" style="color:#072751" class="show" align="center">Welcome to the 2014 GDS iGEM Wiki</p>
<H1 id="mainhead" style="color:#072751" class="show" align="center">Welcome to the 2014 GDS iGEM Wiki</p>
 +
<h2 style="color:#072751">>Projects>CadmiumDetector</h2>
</div>
</div>
-
               
+
<div class="grid-pad" align="center" id="navbar">
<div class="grid-pad" align="center" id="navbar">
<div class="col-1-9">
<div class="col-1-9">
-
<a href="https://2014.igem.org/Team:Gaston_Day_School#home"><p class="currentpage" id="pad">Home<p></a>
+
<a href="https://2014.igem.org/Team:Gaston_Day_School#home"><p class="show" id="pad">Home<p></a>
</div>
</div>
<div class="col-1-9">
<div class="col-1-9">
Line 285: Line 283:
<p class="show" id="pad">Safety<p></a>
<p class="show" id="pad">Safety<p></a>
</div>
</div>
-
<div class="col-1-9">
+
<div class="col-1-9"><a href="https://2014.igem.org/Team:Gaston_Day_School/Attributions#attrib">
-
<a href="https://2014.igem.org/Team:Gaston_Day_School/Attributions#attrib">
+
<p class="show" id="pad">Attributions<p></a>
<p class="show" id="pad">Attributions<p></a>
</div>
</div>
Line 303: Line 300:
<div class="col-1-1">
<div class="col-1-1">
<div class="show" id="pad">
<div class="show" id="pad">
-
<h2 id="head">School Info</h2>
+
<h2 id="head">Cadmium Detector Project Overview</h2>
<p id="pad">
<p id="pad">
-
                            Founded in 1967, Gaston Day School is a non-sectarian, college preparatory institution for grades Pre-K through 12. Throughout its history, nearly 100% of Gaston Day School's classes have entered four-year colleges and universities immediately upon graduation. Located in Gastonia, North Carolina, the school serves communities in a five-county area: Gaston, Lincoln, Cleveland, Mecklenburg and York counties. Enrollment is approximately 500 students. Gaston Day School is accredited by the Southern Association of Colleges and Schools. The school is a member of the Southern Association of Independent Schools, the National Association of Independent Schools, and the North Carolina Association of Independent Schools.The students on the team are in grade 10-12, and each has completed (or is currently in) Honors Biology, Honors Chemistry, Honors Physics, AP Biology, AP Chemistry, AP Physics or some combination thereof.
+
The surrounding areas of Duke Energy’s Buck Steam Station have unintentionally been affected with millions of tons of coal ash containing multiple toxic chemicals including Cadmium. The release of this ash has caused the water to become a health hazard with the potential to cause a wide range of symptoms: flu-like symptoms, kidney damage, fragile bones, and possibly death through prolonged exposure. To minimize the damage caused by Cadmium in water both locally and globally, our 2012 team created several heavy metal detectors, but in 2013 we decided to concentrate on the Cadmium detector. The detector responds to the presence of Cadmium with green fluorescence. In 2014, the team worked to increase the sensitivity levels of our detector. Our detector needed to be able to respond to Cadmium at low enough levels that the detection would be useful and the presence of cadmium would not already be apparent. This year, we completed the addition of the 2007 Cambridge team's sensitivity tuners to our detector. The sensitivity tuners amplify the signal received by the detector. At the end of last year, after adding the sensitivity tuners, we began to see indications of a peak at lower levels of Cadmium than we had previously thought. To define that peak, we used test points that were closer together. We discovered a peak of fluorescence and identified our detection points.  
-
                        </p>
+
                                                </p>
-
</div>
+
                                        </div>
</div>
</div>
</div>
</div>
-
           
+
</div>
-
            <div class="grid-pad" align="center"><div class="col-1-1"></div></div>
+
 
-
           
+
-
            <div class="grid-pad">
+
<div class="col-1-1">
-
                <div class="col-1-2">
+
<h2 id="head">Cadmium Detector Construction</h2>
-
                    <a href="https://2014.igem.org/Team:Gaston_Day_School/biofuels#biofuels">
+
<div class="show" id="pad">
-
                        <h2 id="head">Biofuels</h2>
+
                         <img src="https://static.igem.org/mediawiki/2014/thumb/6/60/Cd_construction.jpg/800px-Cd_construction.jpg" alt="Cadmium methods" style="width:100%">
-
                    </a>
+
                     </div>
-
                    <p class="show" id="pad" style="height:400px">
+
</div>
-
                         The rising cost of petroleum, coupled with the environmental concerns that go with its use, have led to a recent increase in biofuel research.  With this research has come developments in using bacteria to produce biofuels such as isobutanol and ethanol. These alcohols are favored because they can easily be swapped into our current infrastructure of car and truck engines. At Gaston Day School, we have decided to launch a biofuel-focused project. To create the alcohols, we are developing alcohol-resistant strains of E. coli through artificial selection. Also, we are using PCR to amplify and ligate the genes GlmZ, GlmY, and IlmV, which are used in native alcohol production. The combination of these genes and the alcohol resistant strains are the first steps in our new biofuels project.
+
                        </p>
-
                    </p>
+
<div class="col-1-10">
-
                </div>
+
-
                <div class="col-1-2">
+
-
                    <a href="https://2014.igem.org/Team:Gaston_Day_School/cadmiumdetector#cadmiumdetector">
+
-
                        <h2 id="head">Cadmium Detector</h2>
+
-
                    </a>
+
-
                    <p class="show" id="pad" style="height:400px">
+
-
                        The surrounding areas of Duke Energy’s Buck Steam Station have unintentionally been affected with millions of tons of coal ash, containing multiple toxic chemicals including Cadmium.  The release of this ash has caused the water to become a high health hazard with the potential to cause a wide range symptoms, from flu-like symptoms to kidney damage, to fragile bones and possibly death, getting progressively worse through prolonged exposure.  As the danger worsens, more lives are affected.
+
-
In order to minimize the damage caused by cadmium in water, whether locally or globally, our 2012 team created several heavy metal detectors, but in 2013 decided to concentrate on the cadmium detector. The detector responds to the presence of cadmium with green fluorescence. In 2013, the team worked to increase the sensitivity levels of our detector. Our detector needed to be able to respond to cadmium at low enough levels that the detection would be useful, and the presence of cadmium would not already be apparent.This year, we completed the addition of the 2007 Cambridge team's sensitivity tuners to our detector. The sensitivity tuners amplify the signal received by the detector. At the end of last year, after adding the sensitivity tuners, we began to see indications of a peak at lower levels of cadmium than we had previously thought, so to define that peak, we used test points that were closer together. We discovered a peak of fluorescence, and identified our detection points.
+
-
                     </p>
+
-
                </div>
+
-
            </div>
+
-
        </div>
+
-
       
+
-
<div class="col-1-10">
+
<p class="hide" id="pad"></p>
<p class="hide" id="pad"></p>
</div>
</div>
 +
<div class="col-8-10">
 +
<div class="grid-pad">
 +
<div class="col-1-1">
 +
<div class="show" id="pad">
 +
<h2 id="head">Methods</h2>
 +
<p id="pad">
 +
      <br>Starting Sensitivity Tests
 +
<br>Determine which samples you will run
 +
<br>Find both the positive and negative control samples
 +
<br>Obtain 6 tubes for each sample
 +
<br>Add 4 mL of LB (and appropriate antibiotic for the sample)
 +
<br>Add the following amounts of 1 M Cadmium Chloride to the respective tube
 +
<br>1 = 0 mM  = 0 ul
 +
<br>2 = 1 mM  = 4.1 ul
 +
<br>3 = 5 mM  = 20.5 ul 
 +
<br>4 = 10 mM = 41 ul
 +
<br>5 = 50 mM = 205 ul
 +
<br>6 = .1 M      = 410 ul
 +
<br>Add 100ul of the appropriate cells to each tube
 +
<br>Mix well by inverting / vortexing
 +
<br>Grow overnight
 +
<br>
 +
<br>
 +
<br>Reading Sensitivity Results
 +
<br>Spin cells down for 5’ in clinical centrifuge
 +
<br>Decant supernatant
 +
<br>Resuspend in 1 mL of 1x PBS
 +
<br>Open Logger Pro
 +
<br>Plug in SpectroVis
 +
<br>Fill one cuvette with PBS
 +
<br>Click “Experiment” “Calibrate”Spec. 1
 +
<br>DO NOT SKIP WARMUP
 +
<br>Place “0” in SpectroVis
 +
<br>Click “Finish Calibration” ”Okay”
 +
<br>Click on the rainbow mountain
 +
<br>Click “Absorbance vs. Concentration”
 +
<br>Select 600.9 nm
 +
<br>Click “Okay”
 +
<br>Take an OD 600 reading for each sample
 +
<br>CHANGE TO FLUORESCENCE
 +
<br>Experiment  Change Units  Spec. 1  Flu. 405
 +
<br>Click rainbow mountain
 +
<br>Change wavelength to 380.1 nm
 +
<br>Measure a fluorescence reading for each sample
 +
<br>If the reading is above 1:
 +
<br>Take 100 ul of the sample and place in a clean cuvette
 +
<br>Add 900 ul of 1x PBS
 +
<br>Repeat until reading is below 1
 +
<br>Record the number of dilutions
 +
<br>Calculate the “new” flu. reading
 +
<br>Divide the fluorescence reading by its OD 600 reading and record answer as Flu. / OD
 +
<br>Graph and analyze
 +
 +
 +
                            <div class="col-8-10">
 +
<div class="grid-pad">
 +
<div class="col-8-10">
 +
<div class="grid-pad">
 +
<div class="col-1-1">
 +
<h2 id="head">Cadmium Detector Results</h2>
 +
<div class="show" id="pad">
 +
                        <img src="https://static.igem.org/mediawiki/2014/d/db/Cadmium_graph.jpg" alt="cadmium results" style="width:100%">
 +
                    </div>
 +
</div>
</body>
</body>
</html>
</html>

Latest revision as of 03:43, 18 October 2014

Welcome to the 2014 GDS iGEM Wiki

>Projects>CadmiumDetector

The surrounding areas of Duke Energy’s Buck Steam Station have unintentionally been affected with millions of tons of coal ash containing multiple toxic chemicals including Cadmium. The release of this ash has caused the water to become a health hazard with the potential to cause a wide range of symptoms: flu-like symptoms, kidney damage, fragile bones, and possibly death through prolonged exposure. To minimize the damage caused by Cadmium in water both locally and globally, our 2012 team created several heavy metal detectors, but in 2013 we decided to concentrate on the Cadmium detector. The detector responds to the presence of Cadmium with green fluorescence. In 2014, the team worked to increase the sensitivity levels of our detector. Our detector needed to be able to respond to Cadmium at low enough levels that the detection would be useful and the presence of cadmium would not already be apparent. This year, we completed the addition of the 2007 Cambridge team's sensitivity tuners to our detector. The sensitivity tuners amplify the signal received by the detector. At the end of last year, after adding the sensitivity tuners, we began to see indications of a peak at lower levels of Cadmium than we had previously thought. To define that peak, we used test points that were closer together. We discovered a peak of fluorescence and identified our detection points.

Cadmium methods


Starting Sensitivity Tests
Determine which samples you will run
Find both the positive and negative control samples
Obtain 6 tubes for each sample
Add 4 mL of LB (and appropriate antibiotic for the sample)
Add the following amounts of 1 M Cadmium Chloride to the respective tube
1 = 0 mM = 0 ul
2 = 1 mM = 4.1 ul
3 = 5 mM = 20.5 ul
4 = 10 mM = 41 ul
5 = 50 mM = 205 ul
6 = .1 M = 410 ul
Add 100ul of the appropriate cells to each tube
Mix well by inverting / vortexing
Grow overnight


Reading Sensitivity Results
Spin cells down for 5’ in clinical centrifuge
Decant supernatant
Resuspend in 1 mL of 1x PBS
Open Logger Pro
Plug in SpectroVis
Fill one cuvette with PBS
Click “Experiment” “Calibrate”Spec. 1
DO NOT SKIP WARMUP
Place “0” in SpectroVis
Click “Finish Calibration” ”Okay”
Click on the rainbow mountain
Click “Absorbance vs. Concentration”
Select 600.9 nm
Click “Okay”
Take an OD 600 reading for each sample
CHANGE TO FLUORESCENCE
Experiment  Change Units  Spec. 1  Flu. 405
Click rainbow mountain
Change wavelength to 380.1 nm
Measure a fluorescence reading for each sample
If the reading is above 1:
Take 100 ul of the sample and place in a clean cuvette
Add 900 ul of 1x PBS
Repeat until reading is below 1
Record the number of dilutions
Calculate the “new” flu. reading
Divide the fluorescence reading by its OD 600 reading and record answer as Flu. / OD
Graph and analyze

cadmium results

Retrieved from "http://2014.igem.org/Team:Gaston_Day_School/cadmiumdetector"