Difference between revisions of "Team:MIT/Experiments/miRNA/more experiments"
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<p> We want to understand the sensitivity of our miRNA target site constructs.</p> | <p> We want to understand the sensitivity of our miRNA target site constructs.</p> | ||
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| − | <h2 style="text-decoration:underline"> Set Up </h2> | + | <h2 style="color:#000000;text-decoration:underline"> Set Up </h2> |
<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> | <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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Revision as of 18:49, 16 October 2016
Sensitivity of miRNA Target Sites
Purpose
We want to understand the sensitivity of our miRNA target site constructs.
Set Up
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.
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
