Methods to Increase the Sensitivity of High Resolution Melting Single Nucleotide Polymorphism Genotyping in Malaria

Rachel Daniels; Elizabeth J. Hamilton; Katelyn Durfee; Daouda Ndiaye; Dyann F. Wirth; Daniel L. Hartl; Sarah K. Volkman

doi:10.3791/52839

Journal

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Immunology and Infection

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Methods to Increase the Sensitivity of High Resolution Melting Single Nucleotide Polymorphism Genotyping in Malaria

JoVE Journal
Immunology and Infection

This content is Free Access.

JoVE Journal Immunology and Infection

Methods to Increase the Sensitivity of High Resolution Melting Single Nucleotide Polymorphism Genotyping in Malaria

11,498 Views

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10:27 min

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November 10, 2015

DOI:

10.3791/52839-v

DOI:

10.3791/52839-v

•

10:27 min

November 10, 2015

•

2 Views

Rachel Daniels^1,2, Elizabeth J. Hamilton², Katelyn Durfee², Daouda Ndiaye³, Dyann F. Wirth^2,5, Daniel L. Hartl¹, Sarah K. Volkman^2,4

¹Department of Organismic and Evolutionary Biology,Harvard University, ²Department of Immunology and Infectious Diseases,Harvard T.H. Chan School of Public Health, ³Faculty of Medicine and Pharmacy,Cheikh Anta Diop University, ⁴School of Nursing and Health Sciences,Simmons College, ⁵Institute of Infectious Diseases,Broad Institute

Summary

While high resolution melting analysis offers the ability to differentiate between single nucleotide polymorphisms in a heterogeneous population, mutant allele amplification bias can increase its ability to detect alleles present at relatively low percentages within a sample. This protocol describes improvements that improve the sensitivity of high resolution melting analysis.