A New Concept in Preparation of Reference Materials with Heterozygous Genotype for Molecular Diagnostic Purposes

Authors

  • M. Beránek Institute of Clinical Biochemistry and Diagnostics, Faculty of Medicine and Faculty Hospital, Hradec Králové
  • M. Drastíková Institute of Clinical Biochemistry and Diagnostics, Faculty of Medicine and Faculty Hospital, Hradec Králové
  • V. Palička Institute of Clinical Biochemistry and Diagnostics, Faculty of Medicine and Faculty Hospital, Hradec Králové

Keywords:

gene synthesis, reference material, cloning, thrombophilic mutation, commutability

Abstract

Using the technique of enzymatic gene assembly we developed a compound heterozygous reference material for analysis of G20210A mutation in the Factor II and G1691A in the Factor V genes. The prepared clone contained both wild-type and mutant fragments of the genes. The DNA fragments carrying the mutations were inserted into the cloning vector in the inverse orientation relative to the wild-type ones. The clone performance was verified by DNA sequencing and by the allele discrimination real-time PCR methods. Our novel approach makes it possible to prepare a compound heterozygous reference material for any rare variants at biallelic polymorphic sites if analyzed by real-time PCR or other methods.

Downloads

Published

2014-09-15

How to Cite

Beránek, M., Drastíková, M., & Palička, V. (2014). A New Concept in Preparation of Reference Materials with Heterozygous Genotype for Molecular Diagnostic Purposes. Chemické Listy, 108(9), 882–885. Retrieved from http://chemicke-listy.cz/ojs3/index.php/chemicke-listy/article/view/469

Issue

Vol. 108 No. 9 (2014)

Section

Articles

Most read articles by the same author(s)