Team:UNITN-Trento/Notebook/Labposts/09/02

From 2013.igem.org

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{
{
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"date" : "2013-09-10",
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"date" : "2013-09-02",
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"author" : "fabio",
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"author" : "emil",
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"title" : " <html> EFE produced upon blue light illumination!!that’s totally awesome!!</html> ",
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"title" : "50 shades of red(FP)",
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"content" : " <html> in these last days we got our two final complete circuits!! Both my cloning and pedro’s succeeded so now we have a circuit that produces amilCP+EFE with blue light, and a circuit that produces amilGFP+EFE in the dark. We took some gas cromatografic measurements afterr an evaluated induction time with our LED to see if the devices actually produces ethylene. We could only see a really huge peak) ethylene in the sample under blue light with the final circuit with inverter. So the circuit with inverter seems to work, at least usually. We could not observe ethylene with the circuit that I cloned though.</html> ",
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"content" : "<html> I have a new aim, to determine which is the best chassy between 5&alpha; ,10&beta;, TOP10, TB1, JM109, BL21, I'll transform all that one with the same construct pSB1C3 with pLac and mRFP as reporter gene, I'll do the inocula and when the O.D. will reach 0.7 I'll induce with different concentration of IPTG, this could be useful for the modeling page(0.025 mM; 0,05 mM; 0.075 mM; 0.25 mM; 0.5 mM; 0.75 mM; 1 mM; 1.25 mM; 2.5 mM).<br>After that I'll take 1ml sample at every hour for 5 hours.<br>Finally I'll centrifuge every sample in order to replace the LB with PBS then I'll sonicate every sample for 10 seconds and after a new sonication I'll transfer the surnatant in glass cuvettes for the analisys with the fluorometer! The mRFP adsorbs at 584 nm and emitts at 607 nm. This is a real long work and this is my LAST POST!</html>",
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"tags" : "blue_light"
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"tags" : "E.coli strains"
}
}

Revision as of 09:22, 3 October 2013

{ "date" : "2013-09-02", "author" : "emil", "title" : "50 shades of red(FP)", "content" : " I have a new aim, to determine which is the best chassy between 5α ,10β, TOP10, TB1, JM109, BL21, I'll transform all that one with the same construct pSB1C3 with pLac and mRFP as reporter gene, I'll do the inocula and when the O.D. will reach 0.7 I'll induce with different concentration of IPTG, this could be useful for the modeling page(0.025 mM; 0,05 mM; 0.075 mM; 0.25 mM; 0.5 mM; 0.75 mM; 1 mM; 1.25 mM; 2.5 mM).
After that I'll take 1ml sample at every hour for 5 hours.
Finally I'll centrifuge every sample in order to replace the LB with PBS then I'll sonicate every sample for 10 seconds and after a new sonication I'll transfer the surnatant in glass cuvettes for the analisys with the fluorometer! The mRFP adsorbs at 584 nm and emitts at 607 nm. This is a real long work and this is my LAST POST!", "tags" : "E.coli strains" }