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Cat. No.: DN1S-1 (for 1g)

Cat. No.: DN1S-10 (for 10g)

Description

DNase I, also known as Deoxyribonuclease I, is an endonuclease that can digest single-stranded or double-stranded DNA, producing single deoxyribonucleotides or short oligodeoxyribonucleotides. The product of DNase I digestion of single-stranded or double-stranded DNA has a phosphate group at the 5' end and a hydroxyl group at the 3' end.

DNase I activity is dependent on calcium ions and can be activated by magnesium or divalent manganese ions. In the presence of magnesium ions, DNase I can randomly cleave double-stranded DNA at any position. In the presence of divalent manganese ions, DNase I can cleave both strands of DNA at the same position, resulting in blunt ends or sticky ends with 1-2 nucleotides overhang.

Features

RNase-free, suitable for various RNA samples. Provides EDTA for DNase I inactivation when needed.

Applications

Preparation of DNA-free RNA samples; removal of genomic DNA contamination in RNA samples before RT-PCR; removal of DNA templates after in vitro transcription catalyzed by RNA polymerases such as T7, T3, SP6; DNase I footprinting to study DNA-protein interactions; nick translation; generation of random DNA fragment libraries; partial digestion of genomic DNA as a positive control in TUNEL assay for detecting apoptosis.

Source

Purified from bovine pancreas.

Molecular weight

Approximately 32 kDa (monomer).

Activity definition

The amount of enzyme required to completely degrade 1 μg of pBR322 plasmid DNA at 37°C for 10 minutes is defined as one unit of activity.

Activity detection conditions

40 mM Tris-HCl (pH 8.0), 10 mM MgSO4, 1 mM CaCl2, 1 μg of pBR322 DNA.

Purity

Free of other DNA endonucleases and exonucleases, free of RNAse.

Enzyme storage solution

50 mM Tris-acetate (pH 7.5), 10 mM CaCl2, 50% (v/v) glycerol.

Reaction Buffer (10X)

100 mM Tris-HCl (pH 7.5 at 25°C), 25 mM MgCl2, 1 mM CaCl2.

Inactivation or inhibition

DNase I can be inactivated by heating at 65°C for 10 minutes after adding EDTA to a final concentration of 2.5 mM. DNase I can also be inactivated by phenol-chloroform extraction. Chelating agents for metal ions, zinc ions at millimolar concentrations, 0.1% SDS, reducing agents such as DTT and mercaptoethanol, and salt concentrations above 50-100 mM significantly inhibit DNase I.

Storage

Store at -20°C.

Note

• The enzyme should be stored in an icebox or on an ice bath during use, and after use, it should be immediately stored at -20°C.
• This product is for scientific research purposes by professionals only and should not be used for clinical diagnosis or treatment. It should not be used for food or drugs and should not be stored in a regular household.
• For your safety and health, please wear laboratory attire and disposable gloves when handling.

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Only for research and not intended for treatment of humans or animals

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