Part:BBa_K523015:Experience
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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
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