Micrococcal nuclease is derived from Staphylococcus aureus and is a relatively non-specific endo-exonuclease. It is purified from a recombinant E. coli strain that digests double-stranded, single-stranded, circular and linear nucleic acids. The enzyme is active in the pH range of 7.0 - 10.0, with optimal activity at pH 9.2 for both RNA and DNA substrates. Cleavage preferences have been observed at sites rich in adenylate, deoxyadenylate or thymidylate (1). Both DNA and RNA are degraded to 3´ phosphomononucleotides and dinucleotides.
Figure 1: Digestion of 1 µg of Lambda genomic DNA with Micrococcal Nuclease in a 3-fold dilution series. The amount of enzyme used in Lane 2 is defined as 1 gel unit. Lane M is the PCR Marker (NEB #N3234).
Product Source
An E. coli strain containing a genetic fusion of the micrococcal nuclease gene (Gene ID: 3238436) and the gene coding for maltose binding protein, or MBP. The micrococcal nuclease is cleaved from the fusion protein and purified away from MBP.
This product is related to the following categories:
One unit is defined as the amount of enzyme required to digest 1 µg of Lambda DNA in 15 minutes at 37°C, to the extent that the accumulation of low molecular DNA fragments is <400 base pairs as determined by agarose gel electrophoresis.
Reaction Conditions
1X Micrococcal Nuclease Reaction Buffer
Supplement with 100 µg/ml Recombinant Albumin, Molecular Biology Grade
Incubate at 37°C
1X Micrococcal Nuclease Reaction Buffer
50 mM Tris-HCl
5 mM CaCl2
(pH 7.9 @ 25°C)
Storage Buffer
10 mM Tris-HCl
50 mM NaCl
1 mM EDTA
50% Glycerol
pH 7.5 @ 25°C
Heat Inactivation
No
Advantages and Features
Application Features
Degrade nucleic acids present in protein preparations
In vitro translation (2)
Reduce the viscosity of cell lysates during non-mechanic cell lysis preparation
Addition of 1-5 mM Ca++ will increase activity if there is any EDTA or EGTA present
Addition of 1-5 mM EDTA or EGTA will chelate Ca++ and substantially decrease activity
Enzyme inhibited by higher than 100 mM NaCl
Used for chromatin structure analysis (CHiP)
Citations & Technical Literature
Citations
Product Citation Tool
Quality, Safety & Legal
Quality Control Assays
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
Effective May 2, 2023, Recombinant Albumin (#B9200 at 20 mg/ml) replaced Purified BSA (#B9001 at 10 mg/ml).
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]
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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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