Product Overview

DNase I (Deoxyribonuclease I) is an endodeoxyribonuclease capable of digesting single- or double-stranded DNA. It recognizes and cleaves phosphodiester bonds to produce monodeoxynucleotides or single- or double-stranded oligodeoxynucleotides with phosphate groups at the 5'-terminus and hydroxyl groups at the 3'-terminus.

The activity of DNase I depends on Ca²⁺ and can be activated by divalent metal ions such as Mn²⁺ and Zn²⁺. 5 mM Ca²⁺ protects the enzyme from hydrolysis.

In the presence of Mg²⁺, the enzyme can randomly recognize and cleave any site on any strand of DNA.

In the presence of Mn²⁺, both strands of DNA can be simultaneously recognized and cleaved at nearly the same site, forming flat-end DNA fragments or sticky-end DNA fragments with 1–2 protruding nucleotides.

Key Features

High Salt Tolerance

Maintains robust nuclease activity in reaction systems containing 100–200 mM NaCl, enabling efficient DNA removal without additional desalting steps.

High Activity with RNase-Free Assurance

Efficiently degrades residual DNA templates while preserving mRNA integrity, with stringent RNase control to prevent RNA degradation.

Excellent Stability

Exhibits superior thermostability and freeze–thaw stability compared with imported counterparts, supporting reliable performance during routine use and large-scale manufacturing.

Compatible with Long DNA Templates

Efficiently digests residual DNA derived from both standard and long-template IVT reactions, making it suitable for diverse mRNA production workflows.

GMP-Ready and Animal-Origin Free

Manufactured without animal-derived components or β-lactam antibiotics, with rigorous quality control, excellent batch-to-batch consistency, and DMF support available upon request.

Applications

Removal of Residual DNA Following IVT

Following mRNA in vitro transcription (IVT), residual DNA templates can remain in the reaction mixture. DNase I non-specifically hydrolyzes both single- and double-stranded DNA into short oligonucleotides, enabling efficient removal of template DNA and improving the purity of the final mRNA product.

mRNA Manufacturing Workflows

Suitable for DNA template removal during the production of mRNA vaccines, gene therapies, and other RNA therapeutics in both preclinical and GMP manufacturing settings.

Long-Template IVT Applications

Supports efficient digestion of residual DNA from IVT processes involving long templates, including self-amplifying RNA (saRNA) and other extended transcripts.

Performance Data

FAQ

What types of nucleic acids can DNase I digest?

DNase I can digest both single-stranded and double-stranded DNA by cleaving phosphodiester bonds. It does not target RNA under recommended reaction conditions, making it suitable for DNA removal in RNA workflows.

What are the end products generated by DNase I digestion?

DNase I hydrolyzes DNA into monodeoxynucleotides and short oligonucleotides, producing fragments with 5′-phosphate and 3′-hydroxyl termini.

Which metal ions are required for DNase I activity?

DNase I is a Ca²⁺-dependent endonuclease. Calcium ions help stabilize the enzyme and protect it from self-hydrolysis. The enzyme can also be activated by divalent metal ions such as Mg²⁺, Mn²⁺, and Zn²⁺.

How do Mg²⁺ and Mn²⁺ affect DNase I digestion patterns?

In the presence of Mg²⁺, DNase I randomly cleaves DNA at sites along either strand.

In the presence of Mn²⁺, the enzyme tends to cleave both DNA strands at closely adjacent positions, generating blunt-ended fragments or fragments with 1–2 nucleotide overhangs.

Can DNase I be used in mRNA manufacturing workflows?

Yes. DNase I is commonly used after in vitro transcription (IVT) to remove residual DNA templates, improving the purity of mRNA products for both preclinical research and GMP manufacturing applications.

How should DNase I be stored and transported?

The product should be transported under temperatures below 0°C and stored at −25 to −15°C to maintain optimal stability and enzymatic activity.