Difference between revisions of "Part:BBa K2100000:Experience"
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Experiment in MCF-7: | Experiment in MCF-7: | ||
| − | We transfected MCF-7 cells with 250ng of hEF1a-mKate as a transfection marker and 250 ng pERE3-eYFP to examine the promoter's transcriptional activity by observing increases in yellow fluorescence upon cells being induced with | + | We transfected MCF-7 cells with 250ng of hEF1a-mKate as a transfection marker and 250 ng pERE3-eYFP to examine the promoter's transcriptional activity by observing increases in yellow fluorescence upon cells being induced with 5 nM E2. |
https://static.igem.org/mediawiki/2016/f/f5/T--MIT--pERE3MCF7.png | https://static.igem.org/mediawiki/2016/f/f5/T--MIT--pERE3MCF7.png | ||
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The results show a 9 fold difference in yellow fluorescent output between the induced tHESC cells and the uninduced tHESC 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. | The results show a 9 fold difference in yellow fluorescent output between the induced tHESC cells and the uninduced tHESC 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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===User Reviews=== | ===User Reviews=== | ||
Revision as of 23:03, 17 October 2016
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how you used this part and how it worked out.
Applications of BBa_K2100000
We characterized the synthetic ERE3 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 MCF-7:
We transfected MCF-7 cells with 250ng of hEF1a-mKate as a transfection marker and 250 ng pERE3-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 pERE3-eYFP to examine the promoter's transcriptional activity by observing increases in yellow fluorescence upon cells being induced with 50 nM E2.
The results show a 9 fold difference in yellow fluorescent output between the induced tHESC cells and the uninduced tHESC 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.
User Reviews
UNIQ3111ddda477c25ad-partinfo-00000000-QINU UNIQ3111ddda477c25ad-partinfo-00000001-QINU