Proprietary (patent-pending) nonspecific endonuclease with optimal activity in high salt concentration at 4°C to 50°C
Degrades double-stranded and single-stranded DNA and RNA, linear and circular DNA and RNA, and DNA and RNA hybrids
Ideal in bioprocessing and biomanufacturing workflows for nucleic acid removal from recombinant proteins, enzymes, and viruses and for reducing viscosity in lysates
Generates short oligos (as short as 5 nt) with 5´-phosphate and 3´-OH
Up to 2 times more active than other comparable nucleases at 500 mM NaCl/KCl
Binds exceptionally well to cation exchange resins for easy removal
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:
The following reagents are supplied with this product:
NEB #
Component Name
Component #
Stored at (°C)
Amount
Concentration
M0764S
-20
NEBExpress® Salt Active Nuclease
M0764SVIAL
-20
1 x 0.5 ml
100,000 units/ml
Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to release 32 pmol of FAM from FAM-BHQ1 labeled hairpin oligonucleotide in 1 minute at 25°C in a 50 µL reaction in 25 mM Tris-HCl, 500 mM NaCl, 5 mM MgCl2, 0.005% Tween 20 (pH 8.5 @ 25°C).
Storage Buffer
25 mM Tris-HCl
500 mM NaCl
5 mM MgCl2
50% Glycerol
pH 7.5 @ 25°C
Advantages and Features
Features
Proprietary, patent-pending endonuclease with broad nucleic acid activity and optimally active at high salt concentration
Degrades double-stranded and single-stranded DNA and RNA, linear and circular DNA and RNA, and both strands of DNA and RNA hybrids
Generates short oligos (as short as 5 nt) with 5´-phosphate and 3´OH
Up to 2 times more active than other comparable nucleases at 500 mM NaCl
Easy removal from a reaction by capture on cation exchange resin
Reactions are scalable and compatible with high-throughput experiments
Not glycosylated. Recombinant. Detergent-free.
This enzyme is ideal for:
Nucleic acid (DNA and RNA) removal at various steps of a bioprocessing workflow for proteins, viruses, or small molecule purification; nuclease treatment during cell lysis in viral vector (e.g. AAV) manufacturing
Nucleic acid (DNA and RNA) clearance in lysates for improving protein binding to affinity resins or reducing back pressure
Reducing viscosity in cell lysates
Reducing smearing on SDS-PAGE
Product Notes
NEBExpress Salt Active Nuclease can be added directly to a cell extract, a soluble fraction, or a purified sample to degrade DNA and/or RNA.
NEBExpress Salt Active Nuclease is ≥ 50% active in NaCl/KCl 200 -1000 mM, and remains 15% active in Tris buffer, pH 8.5 without salt.
NEBExpress Salt Active Nuclease requires 1-50 mM MgCl2 or 5-100 mM MnCl2.
NEBExpress Salt Active Nuclease is active at temperatures between 4°C and 50°C.
Phosphate-based buffers (including PBS) and SDS are not recommended, as enzyme activity is drastically reduced.
NEBExpress Salt Active Nuclease can be inactivated by the addition of ≥10 mM EDTA, or incubation with 1 mM DTT for 30 min at 55°C, or incubation 1h at 65°C or 20 min at 95°C.
NEBExpress Salt Active Nuclease can be degraded with Proteinase K (NEB #P8107) or Thermolabile Proteinase K (NEB #P8111).
One unit of NEBExpress Salt Active Nuclease will digest 30 µg of calf thymus DNA in 30 min at 37°C in 25 mM Tris-HCl, 500 mM NaCl, 5 mM MgCl2, pH 8.5 reaction buffer.
NEBExpress Salt Active Nuclease is compatible with:
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications & Change Notifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email [email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
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