Genomic DNA Normalization Using Duplex-Specific Nuclease Whole-genome shotgun sequencing (WGS) is a powerful method for studying reference sequences in genomes. It generates different sequence data, which ultimately results in overlapping sequences. The aligned DNA sequences obtained from the overlapping sequence read the assembly into contigs, which can be read through the computer program. Due to the presence of redundant repetitive sequences in small genomes, the WGS method is not applicable1 (cited in 1). Several methods have proposed to eradicate redundancy in plant genomes that depends on the hypermethylation tendency of repetitive sequences. The use of enzymes or the creation of a genomic library could modify the genome, but is limited to limited plant genomes 2-4 (cited in 1). Another method proposed in this article is called "high C0t DNA analysis", based on DNA renaturation kinetics. In this technique, genomic DNA is denatured and slowly annealed, and then hydroxyapatite chromatography is used for dsDNA separation. With the help of detailed knowledge of DNA reassociation kinetics and expertise in spectrophotometry, high C0t DNA analysis can be applied to any genome 5 - 7 (cited in 1). Shagina and others, in 2010, discovered the application of duplex-specific nuclease (DSN) technology to normalization of eukaryotic genomic DNA. It is a simple and effective method, based on hybridization kinetics that excludes the physical separation of ssDNA and dsDNA. After reassociation, the denatured dsDNA contained a repetitive sequence hydrolyzed by DSN and PCR used for the amplification of ssDNA containing low copy molecules8, 9 (cited in 1). The enzyme DSN isolated from Kamchatka crab which is thermostable and specific for dsDNA10...... middle of paper...... preparation of normalized cDNA libraries enriched with full-length sequences. Bioorganic Khim. 31:170-177.10Shagin DA, Rebrikov DV, Kozhemyako VB, Altshuler IM, Shcheglov, Zhulidov PA, Bogdanova EA, Staroverov DB. (2002). A new method for SNP detection using a new specific duplex nuclease from the crab hepatopancreas. Genome Research 12:1935-1942.11Rodrigue S, Malmstrom RR, Berlin AM, Birren BW, Henn MR, and Chisholm SW. (2009). Whole genome amplification and de novo assembly of single bacterial cells. PLoS One 4:e6864.12Cheung F, Haas BJ, Goldberg SM, May GD, Xiao Y, and Town CD. (2006). Sequencing Medicago truncatula expressed tags sequenced using 454 Life Sciences technology. BMC Genomics 7:272.13Lander ES, Linton LM, Birren B, Nusbaum C, Zody MC, Baldwin J, Devon K, Dewar K. (2001). Initial sequencing and analysis of the human genome. Nature 409:860-921.