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Cat. No.: DNIL-1k (for 1KU)

Cat. No.: DNIL-10k (for 10KU)

Description

This product is recombinant DNase I (Deoxyribonuclease I), which is an endonuclease that digests single-stranded or double-stranded DNA. DNase I recognizes and cleaves phosphodiester bonds, producing single deoxynucleotides or single-stranded or double-stranded oligonucleotides with a 5' phosphate group and a 3' hydroxyl group.

The activity of DNase I is dependent on Ca2+ and can be activated by divalent metal ions such as Mg2+ and Mn2+. In the presence of Mg2+, the enzyme randomly recognizes and cuts at any site on either strand of double-stranded DNA. In the presence of Mn2+, it can recognize and cut at nearly identical sites on both strands of DNA, producing blunt ends or DNA fragments with 1-2 nucleotide overhangs.

The product is provided in a lyophilized powder form, which is more convenient for transport and storage. However, once dissolved into a solution, it should be stored at -20°C for long-term storage.

Activity Definition

One unit (U) of activity is defined as the amount of enzyme required to completely degrade 1 μg of pUC19 plasmid DNA in 10 minutes at 37°C.

Source

Yeast recombinant expression

Activity

>3000 Kunitz units/mg

Purity

>95%

Applications

• Removal of genomic DNA contamination from RNA samples.
• Removal of template DNA after in vitro transcription with RNA polymerase.
• Analysis of DNA-protein interactions using DNase I footprinting.
• Used with DNA Polymerase I for nick translation labeling of DNA.
• Construction of DNA random fragment libraries.
• Positive control in apoptosis TUNEL assays by partial cleavage of genomic DNA.

Quality Control

• Protein Purity: Determined by SDS-PAGE gel electrophoresis, with purity not less than 95%.
• RNase Residue Detection: Incubate 1 U DNase I, RNase-free (Lyophilized) with 500 ng total RNA at 37°C for 1 hour, and use agarose gel electrophoresis to verify that more than 90% of the RNA remains intact.

Precautions

• Use RNase-free tubes when handling RNA samples.
• When using this product to remove DNA from RNA samples, add Murine RNase Inhibitor to the reaction at a final concentration of 1 U/μl to protect RNA from degradation.
• DNase I is sensitive to physical denaturation; avoid vigorous shaking during mixing.
• For your safety and health, wear lab coats, disposable gloves, and masks during experimental procedures.

Storage

Store at 4°C.

Comparison

Related:

• DNase I, RNase-free (from Bovine pancreas)
• DNase I, RNase-free (GMP) (from yeast)
• DNase I, RNase-free (Lyophilized) (from yeast)
• DNase I-ST, RNase-free (from yeast)

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

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