Difference between revisions of "Part:BBa K2100005:Experience"
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===Applications of BBa_K2100005=== | ===Applications of BBa_K2100005=== | ||
| + | The MIT iGEM team used the hEF1a promoter to constitutively express fluorescent proteins as transfection markers in our experiments in HEK293, tHESC, MCF-7, and ISH cells. The amount of fluorescent units observed is an indication of the number of plasmids a particular cell uptakes. The graph below shows two single color controls from one of our experiments. We transfected one well of HEK293 cells with hEF1a-mKate and another with hEF1a-eYFP. The cells that uptook the hEF1a-mKate plasmids showed an increase in only the red fluorescent output, while the cells that uptook the hEF1a-eYFP plasmids showed a clear increase in only the yellow fluorescent output. | ||
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| + | https://static.igem.org/mediawiki/parts/c/c8/T--MIT--hEF1a.png | ||
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| + | Here is an image of HEK293 cells transfected with the eYFP florescent gene cascaded with a hEF1a constituitive promoter. We often used yellow florescence as either a reporter for how our promoter responded to induction in cells or as transfection markers for experiments. | ||
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| + | https://static.igem.org/mediawiki/parts/thumb/4/49/T--MIT--khb_eYFPexpression_hek.jpeg/796px-T--MIT--khb_eYFPexpression_hek.jpeg | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 19:04, 21 October 2016
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K2100005
The MIT iGEM team used the hEF1a promoter to constitutively express fluorescent proteins as transfection markers in our experiments in HEK293, tHESC, MCF-7, and ISH cells. The amount of fluorescent units observed is an indication of the number of plasmids a particular cell uptakes. The graph below shows two single color controls from one of our experiments. We transfected one well of HEK293 cells with hEF1a-mKate and another with hEF1a-eYFP. The cells that uptook the hEF1a-mKate plasmids showed an increase in only the red fluorescent output, while the cells that uptook the hEF1a-eYFP plasmids showed a clear increase in only the yellow fluorescent output.
Here is an image of HEK293 cells transfected with the eYFP florescent gene cascaded with a hEF1a constituitive promoter. We often used yellow florescence as either a reporter for how our promoter responded to induction in cells or as transfection markers for experiments.
User Reviews
UNIQ2e548554c955be9a-partinfo-00000000-QINU UNIQ2e548554c955be9a-partinfo-00000001-QINU