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Identifying Mutations by High Resolution Melting in a TILLING Population of Rice
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Genetics
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JoVE Journal Genetics
Identifying Mutations by High Resolution Melting in a TILLING Population of Rice

Identifying Mutations by High Resolution Melting in a TILLING Population of Rice

DOI:
10.3791/59960-v

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06:10 min

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September 02, 2019

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, , , , , ,
1National Key Laboratory of Rice Biology, Zhejiang Provincial Key Laboratory of Crop Genetic Resources, Institute of Crop Science,Zhejiang University, 2Jiaxing Academy of Agricultural Sciences, 3Institute of Nuclear Agricultural Sciences,Zhejiang University, 4The New Countryside Development Institute at Zhejiang University

Chapters

  • 00:04Title
  • 00:25DNA Extraction
  • 01:15PCR Amplification
  • 02:34HRM Scanning and Mutation Confirmation
  • 04:29Results: HRM Curves and Mutation
  • 05:41Conclusion

Summary

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In this article, we present the protocol that is described as high-resolution melting analysis (HRM)-based Target Induced Local Lesions In Genomes (TILLING). This method utilizes fluorescence changes during the melting of the DNA duplex and is suitable for high-throughput screening of both insertion/deletion (Indel) and single base substition (SBS).

Tags

TILLINGRiceHigh Resolution MeltingMutation IdentificationPCRDNA ExtractionSample PreparationHRM AnalysisThermal CyclingMelting Curve Analysis

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