Difference between revisions of "Team:MIT/Experiments/miRNA/more experiments"
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<h2 style="color:#000000;text-decoration:underline"> Flow Cytometry Analysis </h2> | <h2 style="color:#000000;text-decoration:underline"> Flow Cytometry Analysis </h2> | ||
| + | Program Used:<a href=http://bpteague.github.io/cytoflow/>CytoFlo</a> by <a href=https://2016.igem.org/Team:MIT/Team> Brian Teague </a> | ||
<img src = "https://static.igem.org/mediawiki/2016/9/97/T--MIT--transient_transfection.PNG" style = 'padding: 5px'; width: 500px; height = 150px; float: center; border:5px;'> | <img src = "https://static.igem.org/mediawiki/2016/9/97/T--MIT--transient_transfection.PNG" style = 'padding: 5px'; width: 500px; height = 150px; float: center; border:5px;'> | ||
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Revision as of 00:20, 17 October 2016
General Experimental Set-Up and Data Analysis
miRNA Sensor
Read about here
Transient Transfection
Electroporation
Flow Cytometry Analysis
Program Used:CytoFlo by Brian Teague
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
Purpose
Set Up
Results
