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Test using the whole detection circuit and Beta Amyloid to measure circuits response to inputThis test will vary several circuit parameters to find the ideal sensor and characterize total sensor behavior. |
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This will determine the ideal concentration what amounts of Syk-TEVp for each placement of TCS-Gal4VP16 produce the best signalon/noise ratio as well as characterize our circuit for use in other circuitsoff ratio. |
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BCR (CD79A+B (with and without TCS-Gal4VP16), IgM heavy, IgM light), Lyn, Syk-TEVp, tre-mKate(dox reporter circuit), UAS-eYFP(reporter circuit), hef1a:eBFP(transfection marker), (whatever is needed to make dox work). We will be testing all combinations of CD79(A/B)-(stop/TCS-Gal4Vp16). Dox, Beta Amyloid, beta amyloid (if beta amyloid fails, anti IgM can be used to force the receptor into an active state)
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All combinations of plasmids being tested will be prepared and put into dox ladders
First row is control for basal reporter expression.
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The Expression of the reporter should be much higher in Beta Amyloid than in the previous Beta amyloid- testWe should identify what level of Syk-TEVp produces the best on/off ratio. Our data will produce a scatter plot of eYFP (reporter) as a function of mKate (driven by the same promoter as syk). |
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