Related Experiment Video
Updated: Feb 16, 2026

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis
Published on: February 5, 2014
Simple and efficient screening of patients with Fabry disease with high resolution melting
Gabriela Pasqualim1, Bruna Almeida Dos Santos2, Roberto Giugliani3
1Post-Graduation Program on Genetics and Molecular Biology, UFRGS, Porto Alegre, RS 91501-970, Brazil; Gene Therapy Center, Experimental Research Center, Hospital de Clínicas de Porto Alegre, Porto Alegre, RS 90035-903, Brazil.
Background:
Fabry disease (FD [MIM: 301500]) is a disorder caused by mutations in the alpha-galactosidase gene (GLA), which presents great allelic heterogeneity. The development of fast screening methods may reduce costs and length of diagnosis, being particularly important for screening programs of high-risk female patients. Therefore, the purpose of this study was to develop a pre-sequencing genetic screening method based on high resolution melting (HRM) analysis.
Methods:
We performed HRM analysis in one hundred and three individuals, 79 females and 24 males, with a total of 27 different variants in 30 different genotypes. We standardized a protocol using EvaGreen, a release-on-demand dye specific for HRM, added to the PCR reaction. Amplification was performed in a conventional real-time system with HRM capability.
Results:
All genotypes in all amplicons were distinguishable from wild type. In most amplicons it was even possible to differentiate each genotype from the others.
Conclusion:
We developed a simple, fast and highly sensitive HRM based protocol that may facilitate genetic screening of FD.
Related Concept Videos
Phase Transitions: Melting and Freezing
Comparing Intermolecular Forces: Melting Point, Boiling Point, and Miscibility
Temporary attractive forces like dispersion are present in all molecules, whether they are polar or nonpolar. They...
Simple Pendulum
The period of a simple pendulum depends on two factors: its length and the acceleration due to gravity. The period is completely independent of any other factors, such as mass or maximum displacement. For small displacements, a pendulum is...
Simple Trusses
The most basic planar truss is a simple truss with three members arranged in a triangular formation. This triangular truss is inherently stable and rigid due to its geometry, making it an ideal starting point for creating more...
Chromatographic Resolution
The effectiveness of separation can be evaluated by determining the level of separation between two neighboring peaks in a chromatogram, which represents the individual components of a sample.
In chromatography,...
Simple Harmonic Motion

