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    Duplex-specific nuclease

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Duplex-specific nuclease

cat.# EA001; EA002; EA003

- Specific to double-stranded DNA
- Thermostable
- Inhibited by EDTA

Duplex-specific nuclease (DSN) is an enzyme purified from hepatopancreas of Kamchatka crab (Shagin et al., 2002). DSN shows a strong preference for cleaving double-stranded (ds) DNA and DNA in DNA-RNA hybrid duplexes, compared with single-stranded (ss) DNA and RNA. Moreover, the cleavage rate of short, perfectly matched DNA duplexes by this enzyme is considerably higher than that for nonperfectly matched duplexes of the same length. DSN finds use in various applications to isolate single-stranded DNA from complex nucleic acids, for example in cDNA normalization method (Zhulidov et al., 2004; Zhulidov et al., 2005; Bogdanova et al., 2008), for quantitative telomeric overhang determination (Zhao et al., 2008), and for SNP detection (Shagin et al., 2002).

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Duplex-specific nuclease, lyophilized EA001 Thermostable nuclease from Kamchatka crab hepatopancreas with a strong preference for cleaving double-stranded DNA and DNA in DNA-RNA hybrid duplexes 50 U € 350
EA002 100 U € 500
EA003 10 U € 130

DSN was purified from Kamchatka crab hepatopancreas using modified protocol of (Shagin et al., 2002). Lyophilized DSN enzyme must be stored at +4°C before resolution. Other components should be stored at -20°C.

DNAase activity was measured using modified Kunitz assay (Kunitz, 1950), where unit definition was defined as: the amount of DSN added to 50 μg/ml calf thymus DNA that causes an increase of 0.001 absorbance units per minute. Activity assay was performed at 25°C, in 50 mM Tris-HCl buffer, pH 7.15, containing 5 mM MgCl2.

Notice to Purchaser:

Evrogen Nucleic Acid-related Products (the Products) are intended for research use only. The Products are covered by U.S. Pat. # 7,435,794 and other Evrogen Patents and/or Patent applications pending. By use of these Products, you accept the terms and conditions of the applicable Limited Use Label License #002.

MATERIAL SAFETY DATA SHEET INFORMATION

To the best of our knowledge, these products do not require a Material Safety Data Sheet. However, all the properties of these products (and, if applicable, each of their components) have not been thoroughly investigated. Therefore, we recommend that you use gloves and eye protection, and wear a laboratory coat when working with these products.

References:

  • Bogdanova EA, Shagin DA, Lukyanov SA. Normalization of full-length enriched cDNA. Mol Biosyst. 2008; 4 (3):205-12. / pmid: 18437263
  • Kunitz M. Crystalline desoxyribonuclease; digestion of thymus nucleic acid; the kinetics of the reaction. J Gen Physiol. 1950; 33 :363-377. / pmid: 15406374
  • Shagin DA, Rebrikov DV, Kozhemyako VB, Altshuler IM, Shcheglov AS, Zhulidov PA, Bogdanova EA, Staroverov DB, Rasskazov VA, Lukyanov S. A novel method for SNP detection using a new duplex-specific nuclease from crab hepatopancreas. Genome Res. 2002; 12 (12):1935-42. / pmid: 12466298
  • Zhao Y, Hoshiyama H, Shay JW, Wright WE. Quantitative telomeric overhang determination using a double-strand specific nuclease. Nucleic Acids Res. 2008; 36 (3):e14. / pmid: 18073199
  • Zhulidov PA, Bogdanova EA, Shcheglov AS, Shagina IA, Wagner LL, Khazpekov GL, Kozhemyako VV, Lukyanov SA, Shagin DA. A method for the preparation of normalized cDNA libraries enriched with full-length sequences. Bioorg Khim. 2005; 31 (2):186-94. (in Russian)/ pmid: 15889793
  • Zhulidov PA, Bogdanova EA, Shcheglov AS, Vagner LL, Khaspekov GL, Kozhemyako VB, Matz MV, Meleshkevitch E, Moroz LL, Lukyanov SA, Shagin DA. Simple cDNA normalization using kamchatka crab duplex-specific nuclease. Nucleic Acids Res. 2004; 32 (3):e37. / pmid: 14973331
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