Difference between revisions of "Part:BBa K2100002:Experience"
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===Applications of BBa_K2100002=== | ===Applications of BBa_K2100002=== | ||
| − | + | We characterized the synthetic ERE5 promoter in three cell lines: MCF-7, tHESC, and ISH. All cell lines have endogeneous Estrogen Receptor alpha. We analyzed data from cells induced with estradiol (E2) and uninduced as a control. | |
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Experiment in tHESC: | Experiment in tHESC: | ||
Revision as of 23:12, 17 October 2016
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Applications of BBa_K2100002
We characterized the synthetic ERE5 promoter in three cell lines: MCF-7, tHESC, and ISH. All cell lines have endogeneous Estrogen Receptor alpha. We analyzed data from cells induced with estradiol (E2) and uninduced as a control.
Experiment in tHESC:
We transfected tHESC cells with 250ng of hEF1a-mKate as a transfection marker and 250 ng pERE5-eYFP to examine the promoter's transcriptional activity by observing increases in yellow fluorescence upon cells being induced with 5 nM E2.
The results show a fold difference in yellow fluorescent output between the induced MCF-7 cells and the uninduced cells with a vehicle control (0.4% EtOH) to account for the possible increase in proliferation cells undergo when being induced with E2 dissolved in ethanol.
Experiment in tHESC:
We transfected tHESC cells with 250ng of hEF1a-mKate as a transfection marker and 250 ng pERE5-eYFP to examine the promoter's transcriptional activity by observing increases in yellow fluorescence upon cells being induced with 5 nM E2.
The results show a fold difference in yellow fluorescent output between the induced MCF-7 cells and the uninduced cells with a vehicle control (0.4% EtOH) to account for the possible increase in proliferation cells undergo when being induced with E2 dissolved in ethanol.
Experiment in tHESC:
We transfected tHESC cells with 250ng of hEF1a-mKate as a transfection marker and 250 ng pERE5-eYFP to examine the promoter's transcriptional activity by observing increases in yellow fluorescence upon cells being induced with 5 nM E2.
The results show a fold difference in yellow fluorescent output between the induced MCF-7 cells and the uninduced cells with a vehicle control (0.4% EtOH) to account for the possible increase in proliferation cells undergo when being induced with E2 dissolved in ethanol.
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