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

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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;'>

Revision as of 18:50, 16 October 2016

Description of Experiments

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