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Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids
Journal JoVE
Génétique
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Journal JoVE Génétique
Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids

Universal and Efficient Electroporation Protocol for Genetic Engineering of Gastrointestinal Organoids

DOI:
10.3791/60704-v

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07:19 min

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February 18, 2020

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, , , , , ,
1Department of Visceral-, Thoracic and Vascular Surgery, University Hospital Carl Gustav Carus,Technische Universität Dresden

Chapitres

  • 00:05Introduction
  • 00:50Organoid Culture and Preparation
  • 02:23Electroporation
  • 03:33Cell Seeding
  • 04:05Determination of Transfection Efficiency
  • 05:18Results: Electroporation Efficiency of Four Organoid Entities
  • 06:47Conclusion

Summary

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This protocol describes an efficient electroporation method for the transfection of four different gastrointestinal organoid entities with larger plasmids (to the extent of 10 kB). It can be performed within one day and does not need extensive preparation or special, cost-intensive electroporation buffers.

Tags

ElectroporationGenetic EngineeringOrganoids3D Cell CultureTransfectionUniversal ProtocolEfficientTumor OrganoidsHealthy OrganoidsSpheroids2D Cell CultureDissociationY-27632Basal MediumElectroporation BufferPlasmid DNAImpedanceElectroporation Parameters

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