Biopolym. Cell. 2015; 31(4):309-315.
http://dx.doi.org/10.7124/bc.0008F0
Methods
Development of MLPA approach for SNP detection in MTHFR, F5 and F2 genes
1Kravchenko S. A., 1Chernushyn S. Yu., 1Kucherenko A. M., 1Soloviov O. O., 1Livshits L. A.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

Single nucleotide polymorphism (SNP) is an important and valuable form of DNA variation among individuals. Various methods for SNP genotyping have been developed, but a few are suitable for the flexible and low-cost applications. Aim. To design probes and develop diagnostic methods for the analysis of polymorphic variants of the genes MTHFR, F5, F2 using the multiplex ligation-dependent probe amplification (MLPA). Methods. Hybridization and ligation approaches. Results. The specific LPO and RPO probes and primers were designed, the composition and concentration of key components of the mixture for the MLPA reaction, as well as the temperature conditions were selcted. Conclusions. The developed methods are ideally suited for the small-scale SNP genotyping, routinely performed in the medium-sized research laboratories, and can be used for the creation of different test systems for DNA markers.
Keywords: polymorphisms, MLPA, nucleotide mismatch, tautomerisation.
Full text: (PDF, in English)

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