Nuclease P1, Trace Zinc, NaCl Free (Nuclease 5'-Nucleotidehydrolase, 3'-Phosphohydrolase, NP1) available on special order

Nuclease P1 (Nuclease 5'-Nucleotidehydrolase, 3'-Phosphohydrolase) hydrolyzes both 3'-5'- phosphodiester bonds in RNA and heat denatured DNA and 3'-phosphomonoester bonds in mono- and oligonucleotides terminated by 3'-phosphate without base specificity. The enzyme does not attack double-stranded nucleic acids, especially in the presence of more than 400mM of sodium chloride at pH 6.0.\n\nActivity: As reported\n\nUnit Definition:\nOne unit of nuclease activity is defined as the amount of enzyme that produces 1umole of acid soluble nucleotides from RNA based on E = 10,600 for RNA hydrolysates per minute at 37 degrees C, pH 5.3. One unit of the enzyme liberates ~4umoles of orthophosphate from 3пїЅ-AMP per minute at 37 degrees C, pH 7.2.\n\nStable pH: 5.0-8.0 \n\nOptimum pH: 5.3 for RNA and heat-denatured DNA, 7.2 for 3пїЅ-AMP\n\nOptimum Temperature: ~70 degrees C [<60 degrees C for long incubations]\n\nHeat Stability: Stable below 60 degrees C at pH 5.3 for 30 min.\n\nSubstrate Specificity:\nThe enzyme hydrolyzes both 3'-5'- phosphodiester bonds in RNA and heat denatured DNA and 3'-phosphomonoester bonds in mono- and oligonucleotides terminated by 3'-phosphate without base specificity. The enzyme does not digest double-stranded nucleic acids, especially in the presence of more than 400mM of sodium chloride at pH 6.0.\n\nImportant Note: \nN7000-01 does NOT contain sodium chloride or added zinc. Adjustments to previous protocols may be needed.\n\nRecommended Preparation:\nFor maximum recovery of product, briefly centrifuge the original vial prior to opening. This will dislodge any enzyme that has adhered to the side or in the cap due to shipping. Add ddH2O or appropriate buffer. Use a pipet to mix GENTLY the buffer with the nuclease in the vial. Nuclease tends to become layered towards the bottom of the vial. JUST BEFORE ADDITION TO THE REACTION MIXTURE MAKE SURE THE SUSPENSION IS UNIFORM. IF NOT, MIX GENTLY with a pipette BEFORE PIPETTING THE ALIQUOT FOR THE REACTION.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for stable for 2-3 weeks at 4 degrees C and 3-6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.\n\nAlso Available:\nN7000: Nuclease P1 (Nuclease 5'-Nucleotidehydrolase, 3'-Phosphohydrolase)\nN7000-01: Nuclease P1, Trace Zinc, NaCl Free (Nuclease 5'-Nucleotidehydrolase, 3'-Phosphohydrolase)

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SPECIFICATIONS

Catalog Number

N7000-01

Size

100U

Form

Supplied as a lyophilized powder from 30mM sodium acetate buffer, pH 5.3. See reconstitution note below.

References

US Biological application reference: Machida, K. et al., (2006) J. Virology 80:7199-7207. 1. Fujimoto, M., Kuninaka, A., Yoshino, H.: Agricultural Biological Chemistry 38: 777-783 (1974). 2. Schomburg's ENZYME HANDBOOK for nuclease P1 from P. citrinum. 3. Shishido, K., Ando, T., Cold Spring Harbor Monograph Series 14: 155-185 (1982). 4. Lehman, R., The Enzymes (Boyer, ed.) 14: 193-201 (1981). 5. Martin, S.A., et al., Biochim. Biophys. Acta 867: 76-80 (1986).

Alternative Names

EC=3.1.30.1