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DNA Modifying Enzymes and Cloning >
Nucleases >
DNase I (RNase-free) |
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Prices are in US dollars and valid only for US orders.
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- Isolated from a recombinant source
- Supplied with 10X Reaction Buffer
Description:
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.
Source:
A E. Coli strain that carries an MBP fusion clone of Bovine Pancreatic DNase I.
Applications:- Degradation of DNA template in transcription reactions
- Removal of contaminating genomic DNA from RNA samples
- DNase I footprinting
- Nick Translation
Reagents Supplied:
DNase I Reaction Buffer (10X)
Enzyme Properties
Heat Inactivation:
75°C for 10 minutes
Reaction & Storage Conditions
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
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.
Concentration:
2,000 units/ml
Storage Conditions:
10 mM Tris-HCl
2 mM CaCl2
50% Glycerol
pH 7.6 @ 25°C
Storage Temperature:
-20°C
Notes
General notes:- EDTA should be added to a final concentration of 5 mM to protect RNA from being degraded during enzyme inactivation (3).
FAQs
- Will DNase I work in NEB buffers 1-4?
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
RNase Activity:
Incubation of 100 units of DNase I with 10 ug of double-stranded RNA Ladder for 2 hours at 37°C resulted in the same electrophoretic profile as untreated RNA. Incubation of 2 units of DNase I with 10 ug of single-stranded RNA Ladder for 1 hour at 37°C resulted in the same electrophoretic profile as untreated RNA.
References
- Kunitz, M. (1950) J. Gen. Physiol. 33, 349-362.
- Vanecko, S. and laskowski, M. (1961) J. Biol. Chem. 236, 3312-3316.
- Huang, Z. et al. (1996) Biotechniques 20, 1012-1020.
Reagents Sold Separately
DNase I Reaction Buffer
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