Difference between revisions of "Team:William and Mary/Interlab Study"

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                     <p class="h2WM">Interlab Study</p>
 
                     <p class="h2WM">Interlab Study</p>
 
                     <p class="description"></p>
 
                     <p class="description"></p>
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<p>We participated in the Interlab Measurement Study! We created are fluorescent constructs using DNA synthesis and Gibson Assembly. The constructs we measured were J23101 + I13504, J23106 + I13504, and J23117 + I13504. Results are reported below. After we transformed our constructs into cells, we inoculated individual colonies in separate inoculation tubes. We then performed a series of spins and washes to remove the LB media and resuspend the cells in water. Finally we imaged them and obtained fluorescence values using confocal microscopy. In order to analyze the data, we applied a binary definition of what a cell was to our confocal images. We confirmed that the binary definition was picking up as many of the cells on the image as was reasonably possible. Our software determined the average fluorescence measurement for all the pixels inside the binary 'cells' and we reported the average of these measurements for each sample.
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<p>We participated in the Interlab Measurement Study! We created are fluorescent constructs using DNA synthesis and Gibson Assembly. The constructs we measured were J23101 + I13504, J23106 + I13504, and J23117 + I13504.  
 
<br>
 
<br>
 
We assembled the gBlocks using our <a href = "https://2015.igem.org/Team:William_and_Mary/Protocols:Gibson">Gibson assembly protocol</a>, and imaged the constructs using our <a href = "https://2015.igem.org/Team:William_and_Mary/Protocols:Imaging">imaging protocol</a>.</p>
 
We assembled the gBlocks using our <a href = "https://2015.igem.org/Team:William_and_Mary/Protocols:Gibson">Gibson assembly protocol</a>, and imaged the constructs using our <a href = "https://2015.igem.org/Team:William_and_Mary/Protocols:Imaging">imaging protocol</a>.</p>
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<p> We created our fluorescent constructs using DNA synthesis and Gibson Assembly. After we transformed our assembled constructs into cells, we inoculated individual colonies in separate inoculation tubes and grew them in 5mL of LB containing chloramphenicol at 37*C overnight. Cells were imaged following our imaging protocol: link here. Cells were identified using Nikon Elements Software version 7.1 using binary thresholding on the GFP channel. Nikon Elements Software was then used to calculate mean fluorescence per cell and data was exported to Excel for final data analysis.  </p>
 
<p>Our data are shown below:<p>
 
<p>Our data are shown below:<p>
 
<p><img src = "https://static.igem.org/mediawiki/2015/7/77/WMIMPResults.png" width = 700>
 
<p><img src = "https://static.igem.org/mediawiki/2015/7/77/WMIMPResults.png" width = 700>

Revision as of 23:26, 18 September 2015

NOISE - W&M iGEM

Interlab Study

We participated in the Interlab Measurement Study! We created are fluorescent constructs using DNA synthesis and Gibson Assembly. The constructs we measured were J23101 + I13504, J23106 + I13504, and J23117 + I13504.
We assembled the gBlocks using our Gibson assembly protocol, and imaged the constructs using our imaging protocol.

We created our fluorescent constructs using DNA synthesis and Gibson Assembly. After we transformed our assembled constructs into cells, we inoculated individual colonies in separate inoculation tubes and grew them in 5mL of LB containing chloramphenicol at 37*C overnight. Cells were imaged following our imaging protocol: link here. Cells were identified using Nikon Elements Software version 7.1 using binary thresholding on the GFP channel. Nikon Elements Software was then used to calculate mean fluorescence per cell and data was exported to Excel for final data analysis.

Our data are shown below: