Description
NEBExpress Salt Active Nuclease is a proprietary endonuclease (patent-pending) engineered with broad specificity for all forms of DNA and RNA. This salt active nuclease (SAN) has optimal activity in high salt (500 mM NaCl/KCl) and non-specifically cleaves single-stranded, double-stranded, circular and linear DNA, RNA and both strands of DNA:RNA hybrids, to release short oligonucleotides (as short as 5 nucleotides) with 5´-phosphorylated and 3´-hydroxylated ends.
NEBExpress Salt Active Nuclease exhibits greater than 50% activity in salt concentrations from 200-1000 mM, at pH 7.5 to 10 and in presence of 1-50 mM MgCl2, with optimal activity at 500 mM NaCl/KCl. The enzyme functions at a range of temperatures from 4°C through 50°C, and its high isoelectric point (pI) allows for efficient removal by capture on a cation exchange resin.
NEBExpress Salt Active Nuclease is suitable for viscosity reduction and degradation of unwanted DNA and RNA in cell lysates and soluble fractions. The enzyme is compatible with bioprocessing workflows for proteins, viruses, or small molecule purification, and is suitable for nucleic acid digestion in cell lysis steps in viral vector (e.g. adeno-associated (AAV)) production.
Figure 1: NEBExpress Salt Active Nuclease outperforms market-leading nucleases across a range of NaCl concentrations
NEBExpress Salt Active Nuclease is more active than other market-leading nucleases between 250 and 1,000 mM NaCl. The percent activity of each nuclease (normalized by concentration in μg/ml) was determined by measuring the degradation of a fluorescent dsDNA hairpin oligonucleotide at room temperature in the optimal buffer for each nuclease adjusted to a final salt concentration as indicated.
Figure 2: Activity of NEBExpress Salt Active Nuclease on various substrates across a range of NaCl concentrations
NEBExpress Salt Active Nuclease displays peak activity at 300-600 mM NaCl on dsDNA, ssDNA and RNA. The percent activity of NEBExpress Salt Active Nuclease was determined by measuring the degradation of fluorescent oligonucleotide substrates (dsDNA, ssDNA, RNA) at room temperature in 25 mM Tris-HCl, 5 mM MgCl2, pH 8.5 and 0 to 1,000 mM NaCl.
Figure 3: NEBExpress® Salt Active Nuclease outperforms two market-leading nucleases on 3 substrates at 500 mM NaCl
The percent activity of NEBExpress Salt Active Nuclease and two other nucleases (normalized by concentration in μg/ml) was determined by measuring the degradation of fluorescent oligonucleotide substrates (dsDNA, ssDNA, RNA) at room temperature in the optimal buffer for each nuclease adjusted to a final concentration of 500 mM NaCl.
Figure 4: NEBExpress Salt Active Nuclease degrades DNA quickly and efficiently over a range of temperatures
NEBExpress Salt Active Nuclease (1 U) degrades 30 μg of calf thymus DNA in 30 minutes at 37°C and in 60 minutes at 25°C, when tested at 500 mM NaCl pH 8.5.
L=Quick-Load® Purple 1 kb DNA Ladder (NEB #N0552S).
Product Source
An E. coli strain that carries an engineered version of the endA gene from Aliivibrio salmonicida.
- This product is related to the following categories:
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Protein Tools,
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Exonucleases and Non-specific Endonucleases,
- This product can be used in the following applications:
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Protein Purification,
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Protein Analysis Tools,
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Protein Expression