DNase I, (RNase-free) is an endonuclease that nonspecifically cleaves DNA to release di-, tri- and oligonucleotide products with 5´-phosphorylated and 3´-hydroxylated ends (1,2). DNase I acts on single- and double-stranded DNA, chromatin and RNA:DNA hybrids.
Highlights
Isolated from a recombinant source
Supplied with 10X Reaction Buffer
Product Source
An E. Coli strain that carries an MBP fusion clone of Bovine
Advantages and Features
Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme which will completely degrade 1 µg of pBR322 DNA in 10 minutes at 37°C in DNase I Reaction Buffer.
Complete degradation is defined as the reduction of the majority of DNA fragments to tetranucleotides or smaller.
Reaction Conditions
1X DNase I Reaction Buffer
Incubate at 37°C
1X DNase I Reaction Buffer
10 mM Tris-HCl
2.5 mM MgCl2
0.5 mM CaCl2
(pH 7.6 @ 25°C)
Usage Concentration
2,000 units/ml
Storage Buffer
10 mM Tris-HCl
2 mM CaCl2
50% Glycerol
pH 7.6 @ 25°C
Heat Inactivation
75°C for 10 minutes
Related Products
Materials Sold Separately
Product Notes
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EDTA should be added to a final concentration of 5 mM to protect RNA from being degraded during enzyme inactivation (3).
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We do not recommend using NEB's DNase I as a substitute for Monarch DNase I in the RNA isolation workflow when using the Monarch Total RNA Miniprep Kit (NEB #T2010). Monarch DNase I is optimized for use in this workflow, and is available for purchase as a component of the Monarch Total RNA Enzyme Pack, (NEB #T2019).
References
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Kunitz, M. (1950). J. Gen. Physiol . 33, 349-362.
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Vanecko, S. and laskowski, M. (1961). J. Biol. Chem . 236, 3312-3316.
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Huang, Z. et al. (1996). Biotechniques . 20, 1012-1020.
Selection Charts
Selection Charts
Usage Guidelines
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.
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]
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 following link is to a PDF Safety Data Sheet (SDS) that applies to this product to help you use it safely.
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