Difference between revisions of "Team:MIT/Experiments/miRNA/more experiments"

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<h1 style="color:#000000; background-color:#FF9733; -moz-border-radius: 15px; -webkit-border-radius: 15px; padding:15px; text-align: center; font-family: Verdana"> Sensitivity of miRNA Target Sites </h1>   
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<h1 style="color:#000000; background-color:#FF9733; -moz-border-radius: 15px; -webkit-border-radius: 15px; padding:15px; text-align: center"> Sensitivity of miRNA Target Sites </h1>   
 
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<p> this is a test. What are you testing? </p>
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<h2 style="color:#FF9733; text-decoration:underline"> Purpose</h2>
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<h2 style="text-decoration:underline"> Purpose</h2>
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<h2 style="color:#FF9733; text-decoration:underline"> Set-Up</h2>
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<p> We want to understand the sensitivity of our miRNA target site constructs.  </p>
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<a href=https://2016.igem.org/Team:MIT/Experiments/Promoters/Experiment-Details#transient>Transient transfection</a>
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<h2 style="text-decoration:underline”> Set Up </h2>
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<h2 style="color:#FF9733; text-decoration:underline"> Results</h2>
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<p > We chose one miRNA target site, 451a, and designed and ordered siRNA complementary to the target site. We co-transfected tert-immortalized human endometrial stromal cells with our miRNA sensors and varying concentrations of siRNA then measured the red fluorescence output to gauge repression.  </p>
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<img src = "https://static.igem.org/mediawiki/2016/6/6d/T--MIT--sensor.PNG" style = 'padding: 5px'; width: 400px; height = 400px; float: right; border:5px;'>
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<h2 style="text-decoration:underline"> Results </h2>
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<img src = "https://static.igem.org/mediawiki/2016/9/9c/T--MIT--tHESC_siRNA.PNG" style = 'padding: 5px'; width: 400px; height = 400px; float: right; border:5px;'>
 
<img src = "https://static.igem.org/mediawiki/2016/9/9c/T--MIT--tHESC_siRNA.PNG" style = 'padding: 5px'; width: 400px; height = 400px; float: right; border:5px;'>
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When increasing siRNA-451a 0 to 1 nM, there is a 10 fold repression of red fluorescence. Saturation occurs at approximately 10 nM. More experiments would need to be done to understand the responsiveness between 0 to 1 nM.  </p>
 
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<h1 style="color:#000000; background-color:#FF9733; -moz-border-radius: 15px; -webkit-border-radius: 15px; padding:15px; text-align: center; font-family: Verdana"> miRNA Profile of tHESC </h1>
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<h1 style="color:#000000; background-color:#FF9733; -moz-border-radius: 15px; -webkit-border-radius: 15px; padding:15px; text-align: center"> miRNA Profile of tHESC </h1>
 
<img src = "https://static.igem.org/mediawiki/2016/d/dd/T--MIT--tHESCwEstrogen.PNG" style = 'padding: 5px'; width: 400px; height = 400px; float: right; border:5px;'>
 
<img src = "https://static.igem.org/mediawiki/2016/d/dd/T--MIT--tHESCwEstrogen.PNG" style = 'padding: 5px'; width: 400px; height = 400px; float: right; border:5px;'>

Revision as of 18:40, 16 October 2016

Description of Experiments

Sensitivity of miRNA Target Sites


Purpose

We want to understand the sensitivity of our miRNA target site constructs.

Results


When increasing siRNA-451a 0 to 1 nM, there is a 10 fold repression of red fluorescence. Saturation occurs at approximately 10 nM. More experiments would need to be done to understand the responsiveness between 0 to 1 nM.



miRNA Profile of tHESC