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Updated: Jun 1, 2026
Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis
Published on: February 5, 2014
Robust and efficient genotyping of factor V and prothrombin alleles using amplicon high-resolution melting and
Joaquín Royo1, Luis M Muñiz, Gregorio Hueros
1Departamento de Biología Celular y Genética, Universidad de Alcalá, Campus Universitario, Carretera de Madrid-Barcelona km 33.600, 28870 Alcalá de Henares, Madrid, Spain.
Objectives:
Developing robust HRM (amplicon High Resolution Melting) analysis valid for different commercial reaction mixes, using synthetic control DNA samples and the RotorGeneQ (Qiagen) instrument.
Design And Methods:
126 samples were analyzed for the presence of the factor Leiden and the 20210G>A prothrombin alleles. The four alleles were cloned and used to prepare synthetic controls.
Results:
All mutant alleles present in the sample were successfully detected. Genotyping confidence mean was higher than 95%.
Conclusions:
Cost effective HRM genotyping is very reliable using synthetic control DNAs and the RotorGenQ instrument.
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