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Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry
JoVE 杂志
遗传学
This content is Free Access.
JoVE 杂志 遗传学
Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry

Improved Polymerase Chain Reaction-restriction Fragment Length Polymorphism Genotyping of Toxic Pufferfish by Liquid Chromatography/Mass Spectrometry

DOI:
10.3791/54402-v

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09:34 min

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September 20, 2016

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1National Research Institute of Police Science

Chapters

  • 00:05Title
  • 00:45DNA Extraction
  • 02:58PCR and Enzymatic Digestion
  • 04:32Liquid Chromatography/Electrospray Ionization Mass Spectrometry (LC/ESI-MS) Analysis
  • 07:30Results: Representative Results of the Analysis of Raw Muscle from Takifugu rubripes
  • 08:44Conclusion

Summary

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An improved polymerase chain reaction-restriction fragment length polymorphism method for genotyping pufferfish species by liquid chromatography/mass spectrometry is described. A reverse-phase silica monolith column is employed for separating digested amplicons. This method can elucidate the monoisotopic masses of oligonucleotides, which is useful for identifying base composition.

Tags

Puffer Fish GenotypingToxic Puffer FishPCR-RFLPLiquid Chromatography/mass SpectrometryDNA ExtractionUV SpectrophotometryAnalytical ColumnReagent Contamination

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