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

FIGURE 2: Activity of NEBExpress Salt Active Nuclease on various substrates across a range of NaCl concentrations

FIGURE 3: NEBExpress® Salt Active Nuclease outperforms two market-leading nucleases on 3 substrates at 500 mM NaCl

FIGURE 4: NEBExpress Salt Active Nuclease degrades DNA quickly and efficiently over a range of temperatures

Product Source

This product is related to the following categories:

Protein Tools Products,

Exonucleases and Non-specific Endonucleases Products,

This product can be used in the following applications:

Protein Purification,

Protein Analysis Tools,

Protein Expression

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

Properties & Usage

Unit Definition

Storage Buffer

25 mM Tris-HCl
500 mM NaCl
5 mM MgCl₂
50% Glycerol
pH 7.5 @ 25°C

Product Notes

1. 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.
2. 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.
3. NEBExpress Salt Active Nuclease requires 1-50 mM MgCl₂ or 5-100 mM MnCl₂.
4. NEBExpress Salt Active Nuclease is active at temperatures between 4°C and 50°C.
5. Phosphate-based buffers (including PBS) and SDS are not recommended, as enzyme activity is drastically reduced.
6. 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.
7. NEBExpress Salt Active Nuclease can be degraded with Proteinase K (NEB #P8107) or Thermolabile Proteinase K (NEB #P8111).
8. 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.

9. NEBExpress Salt Active Nuclease is compatible with:

   Reagent	Optimal (≥ 80% Active)	Activity >20%
   Tris-HCl/HEPES	25 to 100 mM pH 8.5	>pH 7.5
   NaCl/KCl	300-750 mM	100-300 and 750-1000 mM
   MgCl2	1-50 mM
   MnCl2	5-100 mM
   CaCl2/SDS/Phosphate buffers	Not recommended
   EDTA	≤1 mM
   Triton X-100	≤1%	≤5%
   Tween®20	≤0.1%	≤3%
   DTT	≤1 mM	≤10 mM
   TCEP	≤0.1 mM	≤5 mM
   Urea	≤2 M	<5 M
   Imidazole	<20 mM	<700 mM
   Glycerol	≤5%	<30%
   Protease inhibitor cocktails	with ≤1 mM EDTA

Protocols

1. General Guidance for NEBExpress^® Salt Active Nuclease Usage (NEB #M0764)
2. NEBExpress^® Salt Active Nuclease (NEB #M0764) for nucleic acid clearance in an E. coli soluble fraction
3. Reducing viscosity in an E. coli lysate with NEBExpress^® Salt Active Nuclease (NEB #M0764)

Selection Charts

• Properties of DNase Enzymes

FAQs

1. What are the benefits of working in a high salt buffer?
2. Does NEBExpress^® Salt Active Nuclease (NEB #M0764) have the same efficiency as Serratia marcescens nuclease?
3. How does the unit definition for NEBExpress^® Salt Active Nuclease (NEB #M0764) compare to other salt-active nucleases?
4. Can I use NEBExpress^® Salt Active Nuclease (NEB #M0764) in a low salt buffer?
5. Is NEBExpress^® Salt Active Nuclease (NEB #M0764) compatible with affinity columns such as IMAC?
6. Can I remove NEBExpress^® Salt Active Nuclease (NEB #M0764) from my sample?
7. After use, can I inactivate NEBExpress^® Salt Active Nuclease (NEB #M0764)?
8. How are NEBExpress^® Salt Active Nuclease (NEB #M0764) and DNase I-XT (NEB #M0570) different?
9. What is the average protein concentration of NEBExpress^® Salt Active Nuclease per tube?
10. What is the molecular weight of NEBExpress^® Salt Active Nuclease?
11. Is NEBExpress^® Salt Active Nuclease glycosylated?

Troubleshooting

• Troubleshooting NEBExpress® Salt Active Nuclease (NEB #M0764)

Citations & Technical Literature

Quality Assurance Statement

Specifications

• M0764S_L_v1

Certificate Of Analysis

• M0764S_v1_10260732

Safety DataSheets

---

NEBExpress^® Salt Active Nuclease

• United States
• Portuguese
• Canadian
• United Kingdom
• Chinese
• Dutch
• European
• French
• German
• Italian
• Japanese
• Korean
• Norwegian
• Spanish
• Australia
• New Zealand

Legal and Disclaimers