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Team:Evry/Interlab Study/Used Devices
From 2014.igem.org
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| + | <h2>Used Biobricks and plasmids</h2> | ||
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| + | <h2>Construction way</h2> | ||
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| + | To test the expression of promoters, they are cloned into the PSB1C3 vector followed by the | ||
| + | <a id="igem-link" target="_blank" href="http://parts.igem.org/Part:BBa_E0240" title="Go to iGEM site">BBa_E0240</a>, a GFP generator. Constructions are inserted in E. coli DH5 alpha that grown on LB medium added with the selection agent, chloramphenicol. | ||
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| + | <h2>Analysis protocol</h2> | ||
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| + | Our team decided to test the fluorescence with a | ||
| + | <a id="igem-link" target="_blank" href="http://www.tecan.com/platform/apps/product/index.asp?MenuID=1813&ID=1918&Menu=1&Item=21.2.10.1.1 " title="Go to iGEM site">TECAN infiniteM200</a> in 96 well plates. | ||
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| + | Fluorescence data were analyzed via following formula that come from <a id="igem-link" target="_blank" href="http://www.biomedcentral.com/1752-0509/4/55 " title="Go to iGEM site">2010 De Jong et al </a><i> Experimental and computational validation of models of fluorescent and luminescent reporter genes in bacteria </i>. | ||
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| + | <a href="https://static.igem.org/mediawiki/parts/0/05/BerkiGEM2006-Promoters.jpg" class="image"></a> | ||
| + | <center><img alt="IMAGE" src="https://static.igem.org/mediawiki/2014/8/88/BMC_formlula.emf.jpg" width="300px;float:left;" class="thumbimage"/><br/><center/><br/> | ||
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| + | Equation A: A(t) is the corrected absorbance of DH5 alpha with the functional reporter system, Au(t)the uncorrected absorbance of DH5 alpha with the functional reporter system and Ab(t)the background absorbance corresponding to LB with chloramphenicol medium absorbance.<br> | ||
| + | Equation B: B(t) is the absorbance of the strain carrying the promoterless vector, Iu(t) the uncorrected fluorescence intensity of DH5 alpha with the functional reporter system and Ib(t) the background fluorescence intensity of LB chloramphenicol medium with the DH5 alpha carrying the promoterless vector.<br> | ||
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Revision as of 16:10, 17 October 2014
Interlab study - Used devices
Used Biobricks and plasmids
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Construction way
To test the expression of promoters, they are cloned into the PSB1C3 vector followed by the BBa_E0240, a GFP generator. Constructions are inserted in E. coli DH5 alpha that grown on LB medium added with the selection agent, chloramphenicol.Analysis protocol
Our team decided to test the fluorescence with a TECAN infiniteM200 in 96 well plates.Fluorescence data were analyzed via following formula that come from 2010 De Jong et al Experimental and computational validation of models of fluorescent and luminescent reporter genes in bacteria .
Equation A: A(t) is the corrected absorbance of DH5 alpha with the functional reporter system, Au(t)the uncorrected absorbance of DH5 alpha with the functional reporter system and Ab(t)the background absorbance corresponding to LB with chloramphenicol medium absorbance.
Equation B: B(t) is the absorbance of the strain carrying the promoterless vector, Iu(t) the uncorrected fluorescence intensity of DH5 alpha with the functional reporter system and Ib(t) the background fluorescence intensity of LB chloramphenicol medium with the DH5 alpha carrying the promoterless vector.
Equation B: B(t) is the absorbance of the strain carrying the promoterless vector, Iu(t) the uncorrected fluorescence intensity of DH5 alpha with the functional reporter system and Ib(t) the background fluorescence intensity of LB chloramphenicol medium with the DH5 alpha carrying the promoterless vector.
