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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 Purified BSA
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
Unit Assay Conditions
1X Micrococcal Nuclease Buffer, 0.1 mg/ml BSA and 1 μg Lambda DNA in a total volume of 50 μl.
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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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