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Development and Testing of Species-specific Quantitative PCR Assays for Environmental DNA Applications
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Environment
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JoVE Journal Environment
Development and Testing of Species-specific Quantitative PCR Assays for Environmental DNA Applications

Development and Testing of Species-specific Quantitative PCR Assays for Environmental DNA Applications

DOI:
10.3791/61825-v

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08:54 min

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November 05, 2020

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, , ,
1Columbia Environmental Research Center,U.S. Geological Survey

Chapters

  • 00:04Introduction
  • 01:18Generation of a Sequence Database of Mitochondrial DNA Sequences from Target and Non-target Species of Interest
  • 02:03Assay Design
  • 03:56Assay Screening and Optimization
  • 06:21Results: A. ligamentina qPCR Assay
  • 08:23Conclusion

Summary

Automatic Translation

Automatic Translation

Environmental DNA assays require rigorous design, testing, optimization and validation before the collection of field data can begin. Here, we present a protocol to take users through each step of designing a species-specific, probe-based qPCR assay for the detection and quantification of a target species DNA from environmental samples.

Tags

Keywords: Environmental DNAEDNAQuantitative PCRQPCRSpecies-specific AssayAssay DesignSequence AlignmentNCBIGeneiousMUSCLEPrimer DesignProbe DesignContaminationConservationWildlife Management

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