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Genetic Variant Detection in the CALR gene using High Resolution Melting Analysis
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JoVE Journal Genetica
Genetic Variant Detection in the CALR gene using High Resolution Melting Analysis

Genetic Variant Detection in the CALR gene using High Resolution Melting Analysis

DOI:
10.3791/61642-v

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08:46 min

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August 26, 2020

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, , , , , ,
1Department of Hematology,University Medical Centre Ljubljana, 2Clinical Institute for Genomic Medicine,University Medical Centre Ljubljana, 3Faculty of Pharmacy,University of Ljubljana

Capitoli

  • 00:04Introduction
  • 01:02Fluorescence-based Quantitative Real-time PCR (qPCR) and Post-qPCR Analysis by HRM
  • 05:32Agarose Gel Electrophoresis
  • 06:50Results: Detection of the Genetic Variants in the CALR Gene
  • 08:21Conclusion

Summary

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High resolution melting analysis (HRM) is a sensitive and rapid solution for genetic variant detection. It depends on sequence differences that result in heteroduplexes changing the shape of the melting curve. By combing HRM and agarose gel electrophoresis, different types of genetic variants such as indels can be identified.

Tags

Keywords: Genetic Variant DetectionCALR GeneHigh Resolution Melting AnalysisMyeloproliferative DisordersEssential ThrombocythemiaPrimary MyelofibrosisQPCR HRM Master MixNegative ControlsPositive ControlsNo Template Control (NTC)Optical Adhesive FilmDissociation AnalysisAmplification PlotFluorescence ReadingsPassive Reference

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