Difference between revisions of "Part:BBa K2100059:Experience"

 
 
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how you used this part and how it worked out.
 
how you used this part and how it worked out.
  
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===Applications of BBa_K2100059===
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===Applications of BBa_K2100051===
  
 
===User Reviews===
 
===User Reviews===
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miRNA Profile of tHESC
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Purpose
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To confirm that tHESC under varying conditions (1nm estradiol and no estradiol) would have distinct miRNA profiles.
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Set Up:
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Tert-immortalized human endometrial stromal cells were transfected with our eight candidate miRNA sensors. Twenty-four hours after transfection, half of the samples were induced with 1nM of estradiol . 48 hours later, flow cytometry was performed and red fluorescence from the mKate protein was measured to indicate the amount of repression from endogenous miRNA (Figure 5).Before the finalized analysis, as seen in figure 5, the different miRNA activity detected by the different miRNA sensors was evident by looking at the different cell populations (Figure 4).
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Results:
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Figure 4
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https://static.igem.org/mediawiki/2016/9/91/T--MIT--miRNA_profile_scatter.PNG
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Cells containing either the 142-5p or 21-5p miRNA sensor belonged to two distinct cell populations. 
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Figure 5
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https://static.igem.org/mediawiki/2016/b/b5/T--MIT--miRNA_profile_final.PNG
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Red Relative Fluorescence was normalized using the relative fluorescence of the transfection marker, BFP. The fold difference between estradiol and no estradiol conditional miRNA activity amongst the eight miRNA candidates ranged from 0 to 0.5 fold.

Latest revision as of 22:51, 26 October 2016


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Applications of BBa_K2100051

User Reviews

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miRNA Profile of tHESC Purpose To confirm that tHESC under varying conditions (1nm estradiol and no estradiol) would have distinct miRNA profiles. Set Up: Tert-immortalized human endometrial stromal cells were transfected with our eight candidate miRNA sensors. Twenty-four hours after transfection, half of the samples were induced with 1nM of estradiol . 48 hours later, flow cytometry was performed and red fluorescence from the mKate protein was measured to indicate the amount of repression from endogenous miRNA (Figure 5).Before the finalized analysis, as seen in figure 5, the different miRNA activity detected by the different miRNA sensors was evident by looking at the different cell populations (Figure 4). Results: Figure 4 T--MIT--miRNA_profile_scatter.PNG Cells containing either the 142-5p or 21-5p miRNA sensor belonged to two distinct cell populations. Figure 5 T--MIT--miRNA_profile_final.PNG Red Relative Fluorescence was normalized using the relative fluorescence of the transfection marker, BFP. The fold difference between estradiol and no estradiol conditional miRNA activity amongst the eight miRNA candidates ranged from 0 to 0.5 fold.