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Transgenic Rodent Assay for Quantifying Male Germ Cell Mutant Frequency
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Biologie
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Journal JoVE Biologie
Transgenic Rodent Assay for Quantifying Male Germ Cell Mutant Frequency

Transgenic Rodent Assay for Quantifying Male Germ Cell Mutant Frequency

DOI:
10.3791/51576-v

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14:45 min

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August 06, 2014

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, , , , , ,
1Environmental Health Science and Research Bureau, Health Canada,Environmental Health Centre

Chapitres

  • 00:05Titre
  • 02:30Exposures, Isolation, and Digestion of Cauda Sperm and DNA Extraction
  • 06:45Day Before Assay: Preparation of Agar and Overnight Culture
  • 07:43LacZ Mutation Assay: Sub-culture Cells
  • 08:43LacZ Mutation Assay: Packaging DNA in Lambda Phage Particles and Preparation of Top Agar
  • 10:36LacZ Mutation Assay: Infection of Cells and Counting Plaques
  • 12:40Results: Measurement of Mutant Frequency in Transgenic Mice Cauda Sperm
  • 13:36Conclusion

Summary

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Traduction automatique

De novo mutations in the male germline may contribute to adverse health outcomes in subsequent generations. Here we describe a protocol for the use of a transgenic rodent model for quantifying mutations in male germ cells induced by environmental agents.

Tags

Keywords: Transgenic Rodent AssayMale Germ CellMutant FrequencyDe Novo MutationsGerm Cell MutagenicityIn Vivo AssayOECD Test GuidelineTransgenic VectorReporter GeneSpermatogenic Cell TypesMutagen ExposureCell PopulationSensitivityAnimal Usage

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