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Biochemistry

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JoVE Journal Biochemistry
Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions
 

Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions

Article DOI: 10.3791/58166-v • 09:11 min • October 2nd, 2018
October 2nd, 2018 •
1, 1, 1, 1, 2, 1
1Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán, 2CONACYT, Facultad de Ingeniería Química, Campus de Ciencias Exactas e Ingeniería, Universidad Autónoma de Yucatán

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This protocol describes an efficient methodology for the extraction and quantification of caffeine in cell suspensions of C. arabica L. and an experimental process for evaluating the enzymatic activity of caffeine synthase with the expression level of the gene that encodes this enzyme.

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Caffeine Extraction Enzymatic Activity Gene Expression Caffeine Synthase Plant Cell Suspensions Analytical Chemistry Biochemistry Biotechnology Caffeine Biosynthesis Invitro Plant Models Acetone Lyophilized Cells Vortex Mixer Rotator Silica Gel Plates TLC (thin-layer Chromatography) Mobile Phase Cyclohexane Acetone UV Lamp Densitometry Filter Paper Vacuum Filter Porcelain Mortar Liquid Nitrogen RNA Isolation Reagent Micro Centrifuge Tube Chloroform Isoamyl Alcohol Equilibrated Phenol
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