DNase I, RNase-Free, HC E1018-A

DNase I, RNase-Free, HC

Cat. No./Spec.: E1018-A/1,000 U 

Concentration: 50 U/µL

Product Description 

DNase I (without RNase) is an endonuclease that can cleave both single-stranded and double-stranded DNA. It hydrolyzes phosphodiester bonds, producing single deoxyribonucleotides and oligodeoxyribonucleotides with 5'-phosphate groups and 3'-OH groups.

The activity of this enzyme is strictly dependent on Ca²⁺ and is activated by Mg²⁺ or Mn²⁺ ions. In the presence of Mg²⁺, DNase I cuts each strand of dsDNA in a statistically random and independent manner. When Mn²⁺ is present, the enzyme almost cuts both DNA strands at the same site, resulting in DNA fragments with blunt ends or overhangs of one or a few nucleotides.

Components

Storage Condition & Shelf Life

Store at -20°C.

Source

Recombinant E. coli strain containing the cloned gene encoding bovine DNase I.

Unit Definition

A unit is defined as the amount of enzyme required to completely degrade 1 µg of plasmid DNA within 1 minute at 37°C.

Enzyme activity is determined in the following mixture: 10 mM Tris-HCl (pH 7.5 at 25°C), 2.5 mM MgCl₂, 0.1 mM CaCl₂, and 1 µg of pUC19 DNA.

One unit of DNase I is equivalent to 0.3 Kunitz units.

Features

- Recombinant enzyme

- Purified from a non-animal host, with low levels of endogenous RNase

Scope of Application 

- For the preparation of DNA-free RNA.

- To remove template DNA after in vitro transcription.

- To prepare DNA-free RNA before RT-PCR and RT-qPCR.

- In conjunction with DNA Polymerase I for DNA labeling through nick translation.

- For the study of DNA-protein interactions using DNase I (RNase-free) footprinting.

- To generate a library of randomly sheared DNA insert fragments. Reaction buffer containing Mn²⁺ is used.