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カフェイン抽出、酵素活性、植物細胞懸濁液からのカフェイン合成酵素の遺伝子発現
JoVE Journal
Bioquímica
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JoVE Journal Bioquímica
Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions
DOI:

09:11 min

October 02, 2018

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Capítulos

  • 00:04Título
  • 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

Summary

Tadução automática

このプロトコルは、抽出と成木L の細胞懸濁液中のカフェインの定量のための効率的な方法論と表現レベルのカフェイン合成酵素の酵素活性を評価するための実験的プロセスについて説明します。この酵素を符号化する遺伝子。

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