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Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions
Chapters
- 00:04Title
- 00:28Caffeine Extraction in Cell Suspension of C. arabica L.
- 00:55Conditions for the Assessment of Caffeine by Thin Layer Chromatography (TLC) Denistometry
- 01:29Ribonucleic Acid (RNA) Isolation
- 02:55Incubation of RNA Transcript with Deoxyribonuclease (DNase)
- 03:59Complementary Deoxyribonucleic Acid (cDNA) Synthesis
- 05:03Real-Time Quantitative PCR (qPCR) to Amplify the Gene CCS1
- 05:43Total Protein of Cell Suspensions of C. arabica L.
- 06:26Enzymatic Assay for Caffeine Synthase
- 07:39Results: Enzymatic Activity of Caffeine Synthase Gene Evaluated via qPCR
- 08:30Conclusion
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.
