Difference between revisions of "Part:BBa K325219:Stability"
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#The protocol can be found as [http://2010.igem.org/Team:Cambridge/Notebook/Week11 Experiment 110] on the [http://2010.igem.org/Team:Cambridge Cambridge iGEM 2010 Website]. | #The protocol can be found as [http://2010.igem.org/Team:Cambridge/Notebook/Week11 Experiment 110] on the [http://2010.igem.org/Team:Cambridge Cambridge iGEM 2010 Website]. | ||
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</div>Measured by the [http://2010.igem.org/Team:Cambridge Cambridge iGEM team 2010] | </div>Measured by the [http://2010.igem.org/Team:Cambridge Cambridge iGEM team 2010] | ||
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[http://www.ncbi.nlm.nih.gov/pubmed/18949818 '''[1]:'''] S.M. Marques and J.C.G. Esteves da Silva, (2009) Firefly Bioluminescence: A Mechanistic Approach of Luciferase Catalyzed Reactions,''Life'' '''61''', 6-17. | [http://www.ncbi.nlm.nih.gov/pubmed/18949818 '''[1]:'''] S.M. Marques and J.C.G. Esteves da Silva, (2009) Firefly Bioluminescence: A Mechanistic Approach of Luciferase Catalyzed Reactions,''Life'' '''61''', 6-17. | ||
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[http://www.nature.com/nature/journal/v440/n7082/abs/nature04542.html '''[2]:'''] T. Nakatsu ''et al.'' (2006) Structural Basis for the spectral difference in luciferase bioluminescence, ''Nature'' '''440'''(16), 372-376. | [http://www.nature.com/nature/journal/v440/n7082/abs/nature04542.html '''[2]:'''] T. Nakatsu ''et al.'' (2006) Structural Basis for the spectral difference in luciferase bioluminescence, ''Nature'' '''440'''(16), 372-376. | ||
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[http://www.ncbi.nlm.nih.gov/pubmed/11457857 '''[3]:'''] K. Gomi and N. Kajiyama, (2001) Oxyluciferin, a Luminescence Product of Firefly Luciferase, Is Enzymatically Regenerated into Luciferin, ''The Journal of Biological Chemistry'', '''276'''(39), 36508-36513. | [http://www.ncbi.nlm.nih.gov/pubmed/11457857 '''[3]:'''] K. Gomi and N. Kajiyama, (2001) Oxyluciferin, a Luminescence Product of Firefly Luciferase, Is Enzymatically Regenerated into Luciferin, ''The Journal of Biological Chemistry'', '''276'''(39), 36508-36513. |
Revision as of 11:16, 22 October 2010
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Description
Both the intensity and the spectrum emitted by the luciferase-luciferin reaction has been shown to be higly dependent on the pH of the medium. The main characterisation experiments have been performed in LB Broth at pH 7, so in order to assess this effect cultures with LB and a citrate buffer were prepared (pH = 5.3, pH 6.1 and pH = 7) This page describes the results of these experiments. We used a [http://www.bmglabtech.com/products/microplate-reader/instruments.cfm?product_id=2 FLUOstar OPTIMA] microplate reader to quantify the light output. Protocols and plate reader settings used are given below.
Data
Data | Notes | Date Uploaded |
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Media:BBa_K325219ArabinosetoLight.xls | Raw data from experiment | 21/10/2010 |
Protocol
- The protocol can be found as [http://2010.igem.org/Team:Cambridge/Notebook/Week11 Experiment 110] on the [http://2010.igem.org/Team:Cambridge Cambridge iGEM 2010 Website].
Compatibility
Chassis: Device has been shown to work in Top 10 (Invitrogen)
Plasmids: Device has been shown to work on pSB1C3
References
[http://www.ncbi.nlm.nih.gov/pubmed/18949818 [1]:] S.M. Marques and J.C.G. Esteves da Silva, (2009) Firefly Bioluminescence: A Mechanistic Approach of Luciferase Catalyzed Reactions,Life 61, 6-17.
[http://www.nature.com/nature/journal/v440/n7082/abs/nature04542.html [2]:] T. Nakatsu et al. (2006) Structural Basis for the spectral difference in luciferase bioluminescence, Nature 440(16), 372-376. </div>
[http://www.ncbi.nlm.nih.gov/pubmed/11457857 [3]:] K. Gomi and N. Kajiyama, (2001) Oxyluciferin, a Luminescence Product of Firefly Luciferase, Is Enzymatically Regenerated into Luciferin, The Journal of Biological Chemistry, 276(39), 36508-36513.