Product Class: Other

Micrococcal Nuclease
Gray property icon for NEBU recombinant Gray property icon for incubation temperature 37° Gray property icon for heat inactivation no Gray property icon for rAlbumin

Product Introduction

Mechanism of Micrococcal Nuclease

Micrococcal Nuclease is ideal for:

  • Preparation of double-stranded DNA fragments with 5'-OH and 3'-phosphate
  • Studies of chromatin structure
  • Degradation of nucleic acids in crude cell-free extracts
  • Preparation of rabbit reticulocyte

 

Bioz Badge Exists : True
Catalog # Size Concentration
M0247S 320000 gel units 2000000 gel units/ml

Product Information

Description

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.

Micrococcal Nuclease works well with the SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003 from Cell Signaling Technology. Please follow the link to see product information and protocols on their website.

Figure 1: Digestion with Micrococcal Nuclease

Gel depicting digestion with Micrococcal Nuclease
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:
Chromatin Analysis,
Exonucleases and Non-specific Endonucleases,
RNA Modification,

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

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

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
  • Chromatic structure analysis (3)
  • Rapid RNA sequencing

Product Notes

  1. This enzyme does not work in NEBuffers r1.1-r3.1 or rCutSmart due to the lack of Ca2+. Additional Ca2+ in NEBuffers only shows 10% activity.
  2. 1-5 mM Ca2+ is required for activity.
  3. The enzyme is active in the pH range 7-10, with optimal activity at 9.2, as long as salt concentration is less than 100 mM.
  4. Enzyme can be inactivated by addition of excess EGTA.

References

  1. Cuatrecasas, S.F., and Anfinsen, C.B. (1967). J. Biol. Chem. 242, 1541-1547.
  2. Craig, D. et al. (1992). Nucl. Acids Res. 20, 4957-4987.
  3. O'Neill, L.P. and Turner B.M. (2003). Methods. 31, 76-82.

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Will Micrococcal Nuclease cleave DNA as well as RNA and what type of end is left after digestion?
  2. Does Micrococcal Nuclease work in ChIP sequencing protocols?
  3. Will Micrococcal Nuclease work in rCutSmart Buffer?

Tech Tips

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)

Quality, Safety & Legal

Quality Assurance Statement

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.

Specification Change Notifications

Effective May 2, 2023, Recombinant Albumin (#B9200 at 20 mg/ml) replaced Purified BSA (#B9001 at 10 mg/ml).

Specifications

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]

Certificate Of Analysis

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]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

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 busdev@neb.com.

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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