Team:Heidelberg/Project/miMeasure
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Hela cells transfected with the constructs (described above) are also taken for flow cytometry. 10000 cells are measured. The cells are plotted on a logarithmic scale in relation to EGFP and EBFP2 intensity. Each dot represents one fluorescent cell. The two different coloured dots belong to two different measurements. Both sets of dots make up a line, which shows the correlation of EGFP and EBFP2 very well. The orange dots represent cells, which were transfected with different miMeasure constructs and the miRsAg. The blue dots represent cells, which were also transfected with the different miMeasure constructs, but here the miR-155 is cotransfected, which doesn't have any specificity for the binding sites. The cotransfection with miR-155 is therefore the negative control. If the dots become white, it means the orange and blue ones overlap. So both lines overlap in the negative control, whereas the orange line is going up for the miMeasure construct with the perfect binding site. All the other constructs look like the negative control. | Hela cells transfected with the constructs (described above) are also taken for flow cytometry. 10000 cells are measured. The cells are plotted on a logarithmic scale in relation to EGFP and EBFP2 intensity. Each dot represents one fluorescent cell. The two different coloured dots belong to two different measurements. Both sets of dots make up a line, which shows the correlation of EGFP and EBFP2 very well. The orange dots represent cells, which were transfected with different miMeasure constructs and the miRsAg. The blue dots represent cells, which were also transfected with the different miMeasure constructs, but here the miR-155 is cotransfected, which doesn't have any specificity for the binding sites. The cotransfection with miR-155 is therefore the negative control. If the dots become white, it means the orange and blue ones overlap. So both lines overlap in the negative control, whereas the orange line is going up for the miMeasure construct with the perfect binding site. All the other constructs look like the negative control. | ||
- | [[Image:Flow_miR122.jpg|thumb| | + | [[Image:Flow_miR122.jpg|thumb|610px|center|'''GFP/BFP correlation of single transfected Hela cells according to flow cytometry analysis''' different binding sites for miR-122 cotransfected with miR-122 or with mi-R155, respectively. The orange dots represent the cotransfected cells with miRsAg and the blue dots the cotransfected cells with miR-155. Hela cells were used.]] |
===Analysis of miRaPCR Generated Binding Sites Against a Natural miRNA=== | ===Analysis of miRaPCR Generated Binding Sites Against a Natural miRNA=== |
Revision as of 18:19, 27 October 2010
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