DNase/RNase Mix - BioReagent

Cat. No.: D1528849

GRADE & PURITY BioReagent, DNase I: 1 mg/mL, RNase A: 0.5 mg/mL

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Application

1096

Overview

Deoxyribonuclease I (DNase I) is a non‑specific endonuclease, mostly derived from recombinant *E. coli* strains with an MBP‑fusion clone of bovine‑pancreatic DNase I. DNase I degrades both single‑stranded and double‑stranded DNA by hydrolyzing phosphodiester bonds to generate mononucleotides or oligonucleotides with 5′‑phosphate and 3′‑OH termini. Its activity can be activated by either Mg²⁺ or Mn²⁺, while Ca²⁺ concentration directly modulates enzymatic performance. In the presence of Mg²⁺, random nicks are introduced into each strand of double‑stranded DNA; in the presence of Mn²⁺, double‑stranded DNA undergoes complete cleavage and fragmentation.

Ribonuclease A (RNase A) digests RNA and is widely applied to remove RNA contamination during plasmid and genomic DNA extraction.

DNase/RNase Mix, also known as DNase‑RNase mixed solution, consists of DNase I, RNase A, Tris‑HCl, preservatives and other components. The final concentrations are 1 mg/mL for DNase I and 0.5 mg/mL for RNase A. This reagent is used to eliminate DNA and RNA contamination in research samples.

For research‑use only. Not intended for clinical diagnosis or any other clinical applications.

Procedure (for reference only):

Add an appropriate amount of the enzyme mixture according to experimental requirements to achieve sufficient digestion of DNA and RNA.

Precautions:

1. Avoid reagent contamination and repeated freeze‑thaw cycles.

2. Wear laboratory coat and disposable gloves for personal safety during handling.

3. Use the reagent promptly after opening to prevent compromised experimental performance.

Specifications

Specifications & Purity

BioReagent, DNase I: 1 mg/mL, RNase A: 0.5 mg/mL

Stability And Storage

Store at -20℃ long term (12 months).

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

BioReagent

Product Line

Biological Products

Application Protocols

Item-specific validated protocols, activity units, and recommended working concentrations are Not specified for this item; refer to CoA/Spec Sheet and your laboratory SOPs.

General literature example (for planning only; not item-specific)

• Lysate clarification
  • Prepare cell or tissue lysate in Tris-HCl (20–50 mM, pH 7.5–8.0), 50–150 mM NaCl.
  • Add MgCl2 to 1–5 mM (and 0.5 mM CaCl2 if DNase I-type activity is needed/stabilized), then add nuclease mix at a low test dose.
  • Incubate 15–30 min at 25–37°C with gentle mixing until viscosity decreases.
  • Quench with 10 mM EDTA (for DNase), chill, and proceed to clarification by centrifugation/filtration.
• Removal of contaminating nucleic acids from protein prep
  • Add nuclease mix post-elution if compatible; incubate briefly; remove enzyme by buffer exchange or additional chromatography. Always confirm compatibility with sensitive targets and downstream assays. Include appropriate negative controls to assess background degradation.

Biological Roles

• General biological function (literature; not item-specific)
  • DNases and RNases catalyze the hydrolytic cleavage of phosphodiester bonds in DNA and RNA, respectively. They play roles in nucleic acid turnover, replication quality control, apoptosis (e.g., DNA fragmentation), RNA maturation/processing, and host defense.
• Mechanistic notes
  • Many DNases operate via metal-ion-assisted catalysis, coordinating Mg2+ (and sometimes Ca2+) to activate water for in-line attack on the phosphate.
  • RNase A-type enzymes utilize general acid–base catalysis with two histidines and a lysine, forming a cyclic 2′,3′-phosphate intermediate.
• Research utility
  • Exogenous nuclease addition in vitro simplifies lysate handling, reduces viscosity, and removes nucleic acid contaminants that interfere with chromatographic separations, electrophoresis, and spectrophotometric assays.
• Source and sequence
  • Specific organismal source(s) and sequence composition for the enzymes in this item are not specified; refer to CoA/Spec Sheet. Accordingly, immunogenicity profiles, glycosylation status, and tags (if any) are not defined for this product.

Buffer Applications

This product is an enzyme mixture, not a buffering agent. It does not serve as a primary buffer component.

• Practical note
  • Use in standard biological buffers (e.g., Tris-HCl, HEPES) appropriate for nuclease activity. Maintain near-neutral pH as required by your application (literature). Specific buffer composition for this item is Not specified; refer to CoA/Spec Sheet.

Green Alternatives

• Environmental and safety perspective (general; not item-specific)

  • Enzymatic nucleic acid digestion is typically a greener alternative to chemical hydrolysis using strong acids/bases or oxidants, offering high specificity at mild conditions (aqueous buffers, ambient temperature), reduced hazardous waste, and improved worker safety.

• Comparison (literature-based)

  | Approach | Conditions | Selectivity | Waste/Health considerations | |---|---|---|---| | DNase/RNase enzymatic mix | Aqueous buffer, near-neutral pH, 20–37°C | High for nucleic acids | Low corrosivity; biodegradable proteins | | Acid/base hydrolysis | Strong HCl/NaOH, heat | Nonspecific (damages proteins, materials) | Corrosive, high neutralization burden | | Oxidative methods (e.g., hypochlorite) | Oxidants, ambient | Broad, damages biomolecules | Hazardous oxidants; potential chlorinated byproducts |

• Trade-offs

  • Enzymes can be inhibited by chelators/detergents and may introduce protein contaminants into samples, necessitating post-treatment cleanup.
  • Shelf life and cold-chain requirements (store at −20°C for this item) increase energy use relative to dry chemicals.

• Greener practice tips

  • Use minimal effective doses, optimize buffer to avoid repeat digestions, and aliquot to reduce waste. Implement enzyme inactivation and recovery steps where feasible to minimize residual active enzyme in effluent.

Pharmaceutical Uses

• Context (general; not item-specific)
  • Nucleases can function as process aids in biomanufacturing to reduce host-cell DNA/RNA in recombinant protein, vaccine, and viral vector preparations. They facilitate viscosity reduction and improve clarification/filtration.
• Regulatory considerations
  • When used in GMP settings, lot-specific activity, clearance validation, residual nuclease limits, adventitious agent testing, and endotoxin/bioburden control are critical. This product is designated Research Use Only; it is not cleared for clinical or diagnostic applications.
• Formulation/excipient role
  • Nuclease enzymes are not typical formulation excipients in the final drug product; rather, they are used upstream during purification with validated removal steps (literature best practice).
• Item-specific note
  • No pharmacopeial monograph, residual limits, or GMP status are specified for this item; consult the CoA/Spec Sheet and your quality unit before use in regulated processes.

Physical Properties

• Item-specific physical properties
  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Concentration, formulation buffer, stabilizers, and excipients: Not specified for this item; refer to CoA/Spec Sheet.
  • pH, activity units, and specific activity: Not specified for this item; refer to CoA/Spec Sheet.
  • Melting/boiling points, density, refractive index: Not applicable to protein mixtures in aqueous formulations; Not specified for this item; refer to CoA/Spec Sheet.
• Practical handling properties (general literature context for nucleases; not item-specific)
  • Solubility: Enzymes are typically supplied in aqueous, buffered solutions (often with salts, stabilizers like glycerol) and are water-soluble (literature).
  • Thermal behavior: Proteins generally denature upon heating; many nucleases retain activity near 37°C and lose activity at elevated temperatures depending on formulation (literature).
  • pH tolerance: Many DNase I and RNase A preparations exhibit optimal activity in near-neutral buffers (approx. pH 7–8), but exact ranges depend on formulation (literature).
  • Cofactors: DNase I commonly requires Mg2+ (and is stabilized by Ca2+) for activity; RNase A does not require divalent cations (literature).
• UV/visible characteristics
  • Typical protein UV absorbance maximum at ~280 nm due to aromatic residues (literature); item-specific extinction coefficient and A280-to-concentration conversion are not specified; refer to CoA/Spec Sheet.

Quality & Grades

• Item-specific grade
  • Grade/Purity: BioReagent (as provided). This denotes suitability for general biological research workflows. It does not, by itself, define activity units, nuclease purity profile, host-source, or endotoxin levels for this item; refer to CoA/Spec Sheet.
• What BioReagent typically implies (general guidance)
  • Manufactured for research use with attention to biochemical performance and typical lab contaminants. May be suitable for nucleic acid/protein sample preparation depending on exact specifications.
  • Not certified for clinical or diagnostic use. Research Use Only applies.
• Specification boundaries
  • Activity definitions, specific activity (U/mg), unit definition, residual contaminant nucleases (e.g., absence of protease), endotoxin, bioburden, and stabilizers are Not specified for this item; refer to CoA/Spec Sheet.
• Stabilizers and implications
  • Presence or absence of stabilizers (e.g., glycerol, BSA) is Not specified for this item. Stabilizers can improve freeze–thaw resistance but may interfere with downstream MS or chromatography; verify compatibility with your workflow.
• Documentation
  • For regulated environments, retain the lot-specific CoA and specification sheet for activity units, formulation, and storage recommendations.

Reaction & Applications

• Core functionality (item purpose)
  • A combined DNase/RNase mix enables broad-spectrum digestion of nucleic acids, simplifying workflows where both DNA and RNA must be removed or reduced.
• Typical research applications (literature/practice; not item-specific)
  • Viscosity reduction and clarification of cell lysates prior to protein purification or immunoprecipitation by degrading chromosomal DNA and abundant cellular RNA.
  • Removal of residual host-cell DNA/RNA in protein preparations and enzyme preps.
  • Footprinting/mapping applications where nonspecific nucleic acid removal is required upstream or downstream of the analytical step.
  • Sample cleanup for microscopy or flow cytometry when nucleic acid-associated clumping must be minimized.
  • Processing of crude viral or bacteriophage preparations to remove contaminating nucleic acids outside of capsids.
• Practical tips (general)
  • Add Mg2+ for DNase-dependent steps; Ca2+ can stabilize DNase I. For RNase-driven steps, ensure no RNase inhibitors or strong chelators are present.
  • Titrate enzyme dose to minimize over-digestion of nucleic acid-based reagents (e.g., nucleotide cofactors) and to protect nucleic acid standards if present.
  • Quench by adding EDTA (to inhibit DNase I), by heat denaturation (if compatible with your matrix), or by addition of specific nuclease inhibitors.
  • For downstream mass spectrometry or chromatography, consider potential carryover of proteinaceous enzyme and stabilizers; implement cleanup (e.g., filtration, IMAC flow-through, or protease digestion) as needed.

Reaction Conditions

The following are general, literature-based guidelines for nuclease digestions and not specifications for this item. Optimize empirically for your sample matrix.

• Temperature and time (literature)
  • Many protocols use 25–37°C for 5–60 minutes. Cooler temperatures can preserve sensitive protein complexes at the expense of rate.
• pH and buffers (literature)
  • Near-neutral buffers (Tris-HCl or HEPES, pH ~7.4–8.0) are common. Avoid extreme pH that denatures proteins or nucleic acids before digestion is complete.
• Divalent cations (literature)
  • DNase I-type activity: 1–5 mM MgCl2 is typical; 0.1–1 mM CaCl2 can enhance stability/specificity. EDTA quenches by chelating Mg2+.
  • RNase A-type activity: does not require divalent cations; activity persists in many salt conditions.
• Salt and additives (literature)
  • 50–150 mM NaCl is commonly tolerated. High anionic detergent levels (e.g., ≥0.1% SDS) and strong chaotropes inhibit.
• Enzyme dose (literature)
  • Titrate from low milliunits to a few units per mL of lysate or per mg of total protein, depending on viscosity and nucleic acid load. Start low to avoid over-digestion of nucleic acid-based cofactors.
• Quenching and cleanup (literature)
  • Add EDTA to stop DNase, heat-inactivate if compatible, or remove enzymes via filtration/affinity steps. For RNase, use RNase inhibitors or denaturation/oxidation where appropriate.
• Compatibility
  • For downstream nucleic acid analysis, avoid using this mix; instead, use selective DNase- or RNase-only steps as required.

Safety & Handling

• GHS and hazard data for this specific item
  • Signal Word: Not specified for this item; refer to SDS.
  • H-Statements: Not specified for this item; refer to SDS.
  • GHS Classification and Pictograms: Not specified for this item; refer to SDS.
• General safety notes for nuclease enzymes (literature/practice; not item-specific)
  • Potential allergen/respiratory sensitizer: Avoid generating aerosols or inhalation of protein powders/aerosols. Handle in a biosafety cabinet if aerosolization is possible.
  • PPE: Lab coat, nitrile gloves, and safety glasses. Use eye/face protection when splashing is possible.
  • Incompatibilities: For DNase activity, chelators (e.g., EDTA) and high phosphate can inhibit by sequestering Mg2+; anionic detergents and strong oxidants can denature proteins. RNase inhibitors (protein-based or vanadyl complexes) inhibit RNase activity.
  • Decontamination: Surfaces and tools can be decontaminated with protein denaturants/oxidants (fresh 0.5–1% sodium hypochlorite), strong detergents (SDS), or heat where appropriate. For RNase, standard ethanol is insufficient; use RNase decontamination solutions.
  • First aid (overview): If skin contact occurs, wash with soap and water. If eye contact, rinse cautiously with water for several minutes and seek medical attention if irritation persists. If inhaled, move to fresh air. If ingested, rinse mouth and seek medical advice. Always consult the SDS for authoritative guidance.
• Waste: Collect enzyme-containing waste according to institutional biosafety procedures; deproteinize/inactivate before drain disposal if permitted by local regulations.

Solvent Selection

This product is an aqueous enzyme preparation; organic solvent selection is generally not applicable.

• Applicability
  • Intended for use in aqueous, nuclease-compatible buffers. Organic solvents can denature proteins and are typically avoided.
• Buffer compatibility (general, literature-based; not item-specific)
  • DNase activity typically requires free Mg2+; avoid strong chelators (EDTA, EGTA) during digestion unless intentionally quenching.
  • RNase A family enzymes are broadly active in many neutral buffers; avoid RNase inhibitors if RNA digestion is desired.
  • High concentrations of anionic detergents (SDS) and chaotropes (guanidine, urea) reduce or abolish activity; mild nonionic detergents (e.g., 0.01–0.1% Triton X-100) may be tolerated depending on formulation.
• Practical selection
  • Choose Tris-HCl or HEPES (pH ~7.5–8.0) with NaCl as needed. Add MgCl2 (and CaCl2 for some DNases) for DNA digestion. For RNA digestion, standard neutral buffers without divalent metals are often adequate for RNase A-type enzymes.
• If solvent aspects are central to your assay (e.g., high-organic HPLC mobile phases), perform nuclease digestion in an aqueous step prior to exposure to organic solvents.

Storage & Reconstitution

• Item-specific storage
  • Storage Conditions: Store at -20°C (as provided in Product Data). Avoid repeated freeze–thaw cycles; aliquot upon first thaw when possible.
  • Shipped In: Ice chest + Ice pads (as provided).
• Stability and handling (general guidance; not item-specific)
  • Keep on ice during use. Return promptly to −20°C after aliquoting. If supplied with glycerol, preparations may remain liquid at −20°C and tolerate multiple freeze–thaw events better; formulation for this item is Not specified.
  • Protect from microbial contamination; use sterile, nuclease-free tips and tubes.
• Reconstitution
  • Physical state and need for reconstitution are Not specified for this item; refer to CoA/Spec Sheet.
  • If supplied lyophilized (general case), reconstitute in nuclease-free buffer suitable for your application (e.g., 10–50 mM Tris-HCl, pH 7.5–8.0, with NaCl as needed). Allow to dissolve on ice with gentle mixing; do not vortex vigorously.
• Shelf life
  • Expiry dating and activity retention over time are Not specified for this item; refer to CoA/Spec Sheet.
• Research Use Note
  • For research use only (as provided). Not for diagnostic or therapeutic use.

Structure & Identity

This product is a protein enzyme mixture formulated to hydrolyze both DNA and RNA for laboratory workflows.

• Item-specific identifiers
  • CAS: Not specified for this item; refer to CoA/Spec Sheet.
  • InChIKey: Not applicable for protein mixtures.
  • SMILES: Not applicable for protein mixtures.
  • Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Composition and nature
  • Contains a mix of deoxyribonuclease(s) (DNase) and ribonuclease(s) (RNase) to degrade DNA and RNA, respectively. Exact enzyme identities, sources, and ratios are not specified for this item; refer to CoA/Spec Sheet.
  • Structural class: proteinaceous catalysts composed of amino acids with defined 3D folds; no small-molecule structural representation.
• Literature/general structural features (for context; not specific to this item)
  • DNase I is a Ca2+/Mg2+-dependent endonuclease that cleaves phosphodiester bonds in DNA; typical monomeric enzyme with an active-site histidine/acid-base pair and divalent metal coordination (literature).
  • RNase A family members are small, basic proteins that catalyze phosphodiester cleavage in RNA via a two-histidine, one-lysine catalytic triad (literature).
• 2D description in words
  • As proteins, these enzymes are polypeptide chains forming compact globular structures with surface-exposed basic residues to bind polyanionic nucleic acids, and catalytic pockets that coordinate substrates and, for some DNases, divalent metal ions.

Synthetic Utility

This enzyme mix is not a reagent for small-molecule synthesis. Its value lies in bioprocessing and analytical sample preparation.

• Utility in laboratory workflows (general; not item-specific)
  • Pre-treatment of lysates to reduce viscosity before chromatographic purification of proteins or complexes.
  • Removal of nucleic acids that bind nonspecifically to affinity resins, improving purity and capacity.
  • Clarification of preparations prior to ultracentrifugation or filtration by reducing nucleic-acid-mediated aggregation.
  • Cleanup of PCR setup areas (with appropriate decontamination) by degrading stray nucleic acids; note that RNase presence is incompatible with RNA work unless intentionally applied.
• Retrosynthetic/transform considerations
  • Not applicable to organic retrosynthesis. For nucleic acid biochemistry, controlled digestion can generate defined oligonucleotide fragments for mapping assays (literature).

Target Specificity

Not applicable. This product is a nuclease enzyme mixture, not an antibody or affinity reagent with a molecular target epitope. It non-specifically hydrolyzes nucleic acids (DNA and RNA) according to the enzyme components present.