Russian Journal of Bioorganic Chemistry, Vol. 31, No. 6, 2005, р . 567

Application of the Duplex-Specific Nuclease Preference Method to the Analysis of Single Nucleotide Polymorphisms in Human Genes

I. M. Altshuler*, P. A. Zhulidov**, E. A. Bogdanova**, N. N. Mudrik**, and D. A. Shagin** ^{, 1}

* ZAO Evrogen, Moscow, Russia, ** Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow , 117997 Russia

Abstract: A new modification of the single nucleotide polymorphism (SNP) analysis (DSNP, duplex-specific nuclease preference) method using the duplex-specific nuclease from the king crab was proposed. The method was used to study SNPs in the following human genes: kRAS, nRAS, hRAS, and p53, the genes of blood coagulation factor V, methyltetrahydrofolate reductase, prothrombin, and apolipoprotein E and a deletion in the BRCA1 gene. DSNP was shown to be useful for the estimation of the mutant allele content in DNA samples. A system for the simultaneous identification of several adjacent single-nucleotide polymorphisms in the kRAS gene was proposed. The approaches could be used to develop test systems for the detection of SNPs in human genes.

Key words: DSNP method, duplex-specific nuclease, single-nucleotide polymorphism