title: Robust and efficient genotyping of factor V and prothrombin alleles using amplicon high-resolution melting and synthetic DNA controls: validation of the test with three different commercial chemistries.
abstract: OBJECTIVE
  Developing robust HRM (amplicon High Resolution Melting) analysis valid for different commercial reaction mixes, using synthetic control DNA samples and the RotorGeneQ (Qiagen) instrument.
  
  
  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.
authors.affiliation: Departamento de Biología Celular y Genética, Universidad de Alcalá, Campus Universitario, Carretera de Madrid-Barcelona km 33.600, 28870 Alcalá de Henares, Madrid, Spain.
authors.forename: Joaquín
authors.initials: J
authors.lastname: Royo
authors.forename: Luis M
authors.initials: LM
authors.lastname: Muñiz
authors.forename: Gregorio
authors.initials: G
authors.lastname: Hueros
mesh_terms: D000483:Alleles; D016022:Case-Control Studies; D003001:Cloning, Molecular; D005165:Factor V / Q000235:genetics*; D006579:Heterozygote; D006801:Humans; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D011516:Prothrombin / Q000235:genetics*; D055815:Young Adult
chemical_list: D005165:Factor V; D011516:Prothrombin