Difference between revisions of "Part:BBa K523015:Experience"

 
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===Applications of BBa_K523015===
 
===Applications of BBa_K523015===
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TecCEM_HS 2016
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The TecCEM_HS decided to further work with the part BBa_K523015, which was characterized via a DNS assay. It consists of Lac promoter RBS and endoglucanase from C. Fimi. Optimum temperature was determined.
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Two liters of DNS were prepared with 20 g of NaOH, 4 g of phenol, 1 g of sodium sulfite, 400 g of potassium sodium tartrate and 20 g of DNS. Carboxymethylcellulose (CMC) was used as substrate and all assays were realized with CMC 2% in PBS pH 7. All assays, including the standard curve were done triplicate.
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A standard curve of glucose was realized ranging from 0.1 to 2 mM from a 10 mM stock and diluted with PBS pH 7. In order to reveal the reaction DNS was added and absorbance at 540 nm was determined. Finally data was analyzed in a spreadsheet, plotted obtaining an equation (y=0.8709x-0.1124) with a correlation factor equal to 0.9285
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https://static.igem.org/mediawiki/2016/e/e2/TecCEMHS_StandardCurveDNS.png
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===User Reviews===
 
===User Reviews===

Revision as of 23:19, 20 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_K523015

TecCEM_HS 2016 The TecCEM_HS decided to further work with the part BBa_K523015, which was characterized via a DNS assay. It consists of Lac promoter RBS and endoglucanase from C. Fimi. Optimum temperature was determined.

Two liters of DNS were prepared with 20 g of NaOH, 4 g of phenol, 1 g of sodium sulfite, 400 g of potassium sodium tartrate and 20 g of DNS. Carboxymethylcellulose (CMC) was used as substrate and all assays were realized with CMC 2% in PBS pH 7. All assays, including the standard curve were done triplicate.

A standard curve of glucose was realized ranging from 0.1 to 2 mM from a 10 mM stock and diluted with PBS pH 7. In order to reveal the reaction DNS was added and absorbance at 540 nm was determined. Finally data was analyzed in a spreadsheet, plotted obtaining an equation (y=0.8709x-0.1124) with a correlation factor equal to 0.9285 TecCEMHS_StandardCurveDNS.png


User Reviews

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