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The Zebrafish Tol2 System: A Modular and Flexible Gateway-Based Transgenesis Approach
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Biologia dello sviluppo
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JoVE Journal Biologia dello sviluppo
The Zebrafish Tol2 System: A Modular and Flexible Gateway-Based Transgenesis Approach

The Zebrafish Tol2 System: A Modular and Flexible Gateway-Based Transgenesis Approach

DOI:
10.3791/64679-v

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10:00 min

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November 30, 2022

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, ,
1Alcohol Research Center, Department of Biochemistry and Molecular Genetics,University of Louisville

Capitoli

  • 00:05Introduction
  • 00:53Transposase mRNA Preparation
  • 03:07Multisite Gateway Cloning to Create Entry Vectors for Transgenesis
  • 05:44Injection of the Transgene into the Embryos
  • 07:00Screening Embryos for Transgenic Insertion
  • 07:35Results: Transgenic Constructs of Zebrafish Transgenesis and Their Use in Ethanol Studies
  • 09:34Conclusion

Summary

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This work describes a protocol for the modular Tol2 transgenesis system, a gateway-based cloning method to create and inject transgenic constructs into zebrafish embryos.

Tags

ZebrafishTol2 SystemTransgenesisTransgenicFASD StudiesModularGateway-basedMRNALinearizationEndonucleaseSP6 EnzymeLithium Chloride Precipitation

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