T4 DNA Polymerase

Cat. No.: FP1508928

GRADE & PURITY Suitable for molecular biology, EnzymoPure™, for DNA and RNA applications, 3 U/μL

Overview

  It is purified and prepared from E. coli carrying a highly expressed T4 DNA Polymerase gene.
  T4 DNA Polymerase, namely T4 DNA polymerase, is a template-dependent DNA polymerase, which can catalyze DNA synthesis reaction from the 5'→3' direction on a single-stranded DNA template bound with a primer. T4 DNA Polymerase has 3'→5' exonuclease activity, but no 5'→3' exonuclease activity, and its 3'→5' exonuclease activity is stronger than that of E. coli DNA polymerase I. It is mainly used in the blunting of 5' or 3' protruding ends of DNA, the synthesis of the second strand during site-directed mutagenesis, and the cloning of PCR products independent of ligation reaction, etc.

Component List

FP1508928	Components	300U	3KU	30KU	Storage
FP1508928A	T4 DNA Polymerase (3U/μL)	100 μL	1.0 mL	10 mL	-20℃
FP1508928B	10 ×T4 DNA Polymerase Buffer	1.0 mL	10×1.0 mL	100 mL	-20℃
FP1508928C	Solution I (100×)	100 μL	1.0 mL	10 mL	-20℃

Scope of Application

1. Gap filling (no strand displacement activity) to generate blunt ends;

2. Filling 5' overhangs to form blunt ends;

3. Trimming 3' overhangs to form blunt ends;

4. Probe labeling via displacement synthesis;

5. Subcloning after single-strand deletion;

6. Synthesis of the second strand in site-directed mutagenesis.

Precautions

1. Since this enzyme has 3'→5' exonuclease activity, increasing the reaction temperature, increasing the enzyme amount, not adding dNTPs, or prolonging the reaction time may lead to the excision of bases at the DNA ends, resulting in recessed ends.

2. When using this enzyme, it should be stored in an ice box or on an ice bath, and immediately placed at -20°C for storage after use.

3. For your safety and health, please wear a lab coat and disposable gloves during operation.

Usage Method

  Preparation before reaction system preparation:
a. Dissolve various reagents required for the reaction at 4°C, and place them on an ice bath or in an ice box for use. It is recommended to aliquot the reaction solution for use to avoid repeated freeze-thaw cycles.
b. The reaction system can be set up with reference to the end repair system in the table below, and it is recommended to prepare the system on an ice bath or in an ice box:

Component	Volume	Final Concentration
10×T4 DNA Polymerase Buffer	5 μL	1×
dNTP Mix* (25 mM)	0.4 μL	0.2 mM
Solution I (100×)	0.5 μL	1×
T4 DNA Polymerase (3 U/μl)	—**	—
Reactants	—**	—
ddH₂O	To 50 μL	—
Final Volume	50 μL	—

* 10×T4 DNA Polymerase Buffer does not contain dNTP Mix and needs to be prepared by customers themselves.
** It is recommended to add 1 U of T4 DNA Polymerase per microgram of DNA substrate.

Reaction Procedure (taking end repair as an example)

Step Name	Temperature	Time
Reaction	12~37℃	15~30 min
Inactivation	75℃	20 min
Storage	4℃	∞

Note: The reaction temperature and time can be adjusted according to the specific amount of substrate and enzyme.

Specifications

Specifications & Purity

Suitable for molecular biology,EnzymoPure™,for DNA and RNA applications,3 U/μL

Biochemical and Physiological Mechanisms

Replicates the viral genomic DNA. This polymerase possesses two enzymatic activities: DNA synthesis (polymerase) and an exonucleolytic activity that degrades single-stranded DNA in the 3'- to 5'-direction for proofreading purpose.

Grade

for DNA and RNA applications, Suitable for molecular biology, EnzymoPure™

Application

PCR, qPCR

Storage and Shipping

Storage

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

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at -20℃ long term. Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.

Contents & Storage

FP1508928	Components	300U	3KU	30KU	Storage
FP1508928A	T4 DNA Polymerase (3U/μL)	100 μL	1.0 mL	10 mL	-20℃
FP1508928B	10 ×T4 DNA Polymerase Buffer	1.0 mL	10×1.0 mL	100 mL	-20℃
FP1508928C	Solution I (100×)	100 μL	1.0 mL	10 mL	-20℃

Application Protocols

Item-specific

• No validated protocols, unit definitions, or recommended enzyme concentrations are provided for this item; refer to CoA/Spec Sheet.

Example workflows (literature/general; optimize for your system)

1. Blunt-ending 5′ overhangs (fill-in)

• Mix: DNA (0.1–1 µg), 1× T4 Pol buffer (50 mM Tris-HCl pH 7.8, 10 mM MgCl2, 1 mM DTT), 200 µM each dNTP, and T4 DNA polymerase (per supplier units).
• Incubate: 20 min at 25 °C.
• Stop: Add EDTA to 10 mM, heat 70 °C for 10 min (if compatible), then purify.

2. Blunt-ending 3′ overhangs (chew-back)

• Mix: DNA, 1× buffer as above, omit dNTPs, add enzyme.
• Incubate: 5–15 min at 12–20 °C; monitor to prevent over-digestion.
• Stop and purify as above; proceed to ligation.

3. Second-strand cDNA synthesis (with RNase H)

• After first-strand cDNA synthesis, add RNase H to nick RNA in RNA–DNA hybrid.
• Add T4 DNA polymerase with dNTPs in suitable buffer; incubate 16–25 °C for 30–60 min to extend and replace RNA segments.
• Purify ds cDNA for downstream cloning or library prep.

Controls and QC

• Include no-enzyme negative control and a reference DNA substrate with defined overhangs.
• Validate heat-inactivation and cleanup by test ligation or qPCR amplification.

Biological Roles

Literature/general biochemical role

• Origin: Replicative DNA polymerase from bacteriophage T4 infecting Escherichia coli.
• Function in vivo: Catalyzes leading- and lagging-strand DNA synthesis during phage replication with high fidelity, working in concert with sliding clamp (gp45), clamp loader, primase–helicase complex, and other replisome components.
• Proofreading: Possesses robust 3′→5′ exonuclease activity that excises misincorporated nucleotides, significantly lowering error rates.
• Mechanism: Two-metal-ion catalysis (typically Mg2+) in the polymerase active site; exonuclease active site coordinates metal ions for hydrolytic removal of 3′ nucleotides. DNA shuttles between polymerase and exonuclease sites for error correction.
• Substrate specificity: Requires a primer–template junction (cannot initiate de novo). Accepts standard dNTPs; incorporation fidelity and kinetics are sequence- and condition-dependent. Mn2+ can broaden substrate tolerance but reduces fidelity.
• Biological significance: Serves as a model for family-B polymerases (related to archaeal high-fidelity polymerases). Its strong proofreading explains its utility for laboratory end-polishing and error correction on DNA ends.

Item-specific notes

• No additional biological-source details, expression host, or engineered mutations are provided for this item; refer to CoA/Spec Sheet.

Use limitations

• For research use only; not intended for diagnostic or therapeutic use.

Buffer Applications

Applicability

• Highly applicable. T4 DNA polymerase performance is buffer- and cofactor-dependent. The following are general, literature-based guidelines; this product’s exact buffer system is not specified.

Typical reaction buffer composition (literature)

• 50 mM Tris-HCl, pH 7.5–8.0 at 25 °C
• 5–10 mM MgCl2 (required cofactor)
• 1 mM DTT (reducing environment)
• 50–100 mM NaCl or KCl (ionic strength; optional depending on template)
• 0.05–0.2 mg/mL BSA (stabilizer; optional)
• dNTPs at 50–250 µM each for fill-in reactions; omit to favor 3′ chew-back

Practical buffer tips

• Avoid EDTA carryover from DNA preps; if present, increase Mg2+ accordingly or repurify DNA.
• Glycerol effects: If the enzyme stock contains glycerol, keep final reaction glycerol ≤5–10% to maintain activity; adjust salt accordingly.
• Temperature: Run at 12–37 °C depending on desired balance between exonuclease and polymerase activities (lower temperatures slow exonuclease).
• Quenching: Add 0.1 volume of 0.5 M EDTA or shift to a ligation-ready buffer after cleanup.

Example blunt-ending workflow (literature)

• Assemble DNA (0.1–1 µg), buffer as above, MgCl2, dNTPs (200 µM each), and T4 DNA polymerase (units per manufacturer guidance).
• Incubate 15–30 min at 12–25 °C for controlled polishing.
• Purify DNA and proceed to ligation.

Item-specific

• Exact buffer recipes, unit definitions, and recommended concentrations are not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

Context

• T4 DNA polymerase operates in aqueous buffers at ambient to moderate temperatures. Compared to chemical end-modification methods, enzymatic DNA polishing is inherently greener, avoiding hazardous organic reagents.

Greener practice considerations (literature/general)

• Aqueous-only workflows: Use nuclease-free water and Tris-based buffers; eliminate phenol–chloroform extraction by using silica spin columns or magnetic beads for cleanup.
• Lower energy input: Reactions proceed at ≤37 °C, reducing energy usage relative to high-temperature/denaturing steps.
• Minimize waste: Scale reactions to the minimal volume needed; use concentrated stocks to limit plastic consumables.

Comparison of enzymatic options (trade-offs)

• T4 DNA Polymerase vs Klenow (exo−):
  • Greenness: Both aqueous. Klenow (exo−) minimizes nuclease-generated waste but may leave 3′ overhangs intact; T4 Pol can over-digest if not controlled.
  • Performance: T4 Pol’s strong proofreading is excellent for producing blunt ends but requires tighter time control (risk of material loss).
• T4 DNA Polymerase vs T4 Polynucleotide Kinase + Nucleases:
  • T4 Pol can combine fill-in and chew-back in one step, reducing reagent count.

What is not applicable

• Solvent substitution tables (e.g., replacing THF with 2-MeTHF) are not relevant to this biological reagent.

Operational tips to enhance sustainability

• Use room-temperature-stable master mixes only if validated not to compromise activity.
• Adopt bead-based cleanup to avoid chlorinated solvents and reduce centrifugation time.

Pharmaceutical Uses

Applicability

• Not a pharmaceutical ingredient. This product is intended strictly for research use (as stated in the Product Data) and is not for human or veterinary use.

Relevant non-clinical/industrial contexts (literature/general)

• Biotechnology manufacturing: May be incorporated into in vitro nucleic-acid processing steps during research reagent or diagnostic kit development (e.g., DNA end-polishing modules). Any use in regulated manufacturing would require independent qualification and validation.
• Analytical workflows: Probe generation or DNA labeling for research assays (e.g., Southern blot probes) in quality-control laboratories.
• Companion reagent in device R&D: Used for prototyping of enzymatic DNA assembly or library preparation tools.

What is not claimed here

• No pharmacopeial monograph, excipient role, or clinical application is provided or implied for this item.

Compliance reminder

• Users are responsible for determining suitability, performing method validation, and adhering to applicable regulatory requirements if integrating the enzyme into any GMP or diagnostic workflows.

Physical Properties

Item-specific (from Product Data)

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.

Literature/general properties for T4 DNA polymerase

• Physical form: Typically supplied as an aqueous buffered solution with stabilizers (e.g., glycerol, BSA); exact composition not specified for this item.
• Isoelectric point (pI): Not universally fixed; polymerase family-B pI values often in the mildly acidic to neutral range. Specific pI for this preparation is not specified for this item.
• Temperature profile (literature):
  • Optimal activity around 37 °C for in vitro polymerization on DNA templates.
  • Thermolabile above ~50–60 °C; rapid loss of activity upon heating (e.g., 65 °C heat-inactivation sometimes used but may be incomplete due to strong DNA binding; confirm experimentally).
• pH range (literature): Active in Tris-HCl buffers near neutral to slightly alkaline (pH 7.2–8.4), with Mg2+ as essential cofactor.
• Cofactors: Requires divalent metal (Mg2+ optimal; Mn2+ can be used to alter fidelity/active-site geometry but typically reduces accuracy).
• Solubility: Water-soluble protein in standard molecular biology buffers; insoluble in organic solvents and denatured by SDS and chaotropes.
• Stability influencers: Stabilized by polyols (e.g., glycerol), BSA, reducing agents (DTT); activity inhibited by EDTA (metal chelation), high salt, phenol, and alcohols.

Notes

• Exact density, refractive index, extinction coefficient, and other physicochemical constants are not specified for this item; refer to CoA/Spec Sheet.

Quality & Grades

Item-specific (from Product Data)

• Grade/Purity: Suitable for molecular biology, EnzymoPure™, for DNA and RNA applications.

Interpretation and implications

• Molecular biology grade (general): Indicates the material is manufactured and handled to minimize contaminating activities that compromise nucleic-acid workflows—specifically, free of detectable DNase, RNase, and endonuclease activities under defined test conditions. Low bioburden and low host-nucleic-acid background are typically targeted.
• EnzymoPure™ (general description): A trade descriptor commonly denoting highly purified enzyme suitable for sensitive enzymatic manipulations. While exact specifications are not provided here, such designations often imply:
  • High specific activity and batch-to-batch consistency
  • Stringent QC for contaminating nucleases and proteases
  • Stabilizer system compatible with downstream ligation, PCR, and library-prep steps
• For DNA and RNA applications (item-specific): Indicates compatibility with workflows that contact both DNA and RNA (e.g., cDNA synthesis second-strand, end-polishing of DNA inserts before ligation to RNA adapters), contingent on the absence of RNase contamination.

What is not specified for this item

• Specific activity (U/µL), host expression system, residual host DNA/RNA limits, buffer formulation, and contaminant thresholds are not specified; refer to CoA/Spec Sheet.

Best practices to preserve grade

• Use nuclease-free consumables and buffers; work in a clean area.
• Aliquot to avoid repeated freeze–thaw cycles (see Storage tab).

Reaction & Applications

Item-specific context

• Intended for DNA and RNA applications in molecular biology (from Product Data). Specific unit activity and formulation are not specified; refer to CoA/Spec Sheet.

Key catalytic activities (literature/general)

• 5′→3′ DNA polymerase activity: Extends a primer annealed to template DNA in the presence of dNTPs and Mg2+.
• Strong 3′→5′ exonuclease activity: Proofreads and removes mismatched or recessed 3′ termini, enabling precise end-polishing.
• Lacks 5′→3′ exonuclease activity: Distinguishes it from E. coli Pol I.

Common laboratory applications

• Blunt-end formation: Converts 5′ overhangs to blunt ends by fill-in; converts 3′ overhangs to blunt ends by exonucleolytic chew-back.
• End-polishing before ligation: Prepares DNA fragments for blunt-end ligation or adapter ligation in cloning and NGS library construction.
• Nick translation (with DNase I–generated nicks): Incorporation of labeled nucleotides for probe generation (less common than with Pol I but feasible).
• Second-strand cDNA synthesis: Extends from RNA–DNA hybrids (often in systems where RNase H generates nicks) to produce double-stranded cDNA.
• Site-directed mutagenesis and gap filling: Fills short gaps in DNA repair assays and engineered constructs.

Practical tips

• Control exonuclease chew-back on 3′ overhangs by limiting time, lowering temperature (e.g., 12–25 °C), and maintaining sufficient dNTPs to favor polymerization over degradation.
• Include dNTPs during 5′ overhang fill-in; for pure 3′ chew-back, omit dNTPs but monitor closely to avoid over-digestion.
• Quench with EDTA (10–20 mM final) and/or heat-inactivate as appropriate for your downstream step (confirm heat-inactivation empirically).
• For ligation readiness, purify polished DNA (spin columns or magnetic beads) to remove enzyme, salts, and dNTPs.

Reaction Conditions

Item-specific

• No units, buffer composition, or recommended temperatures are provided for this item; refer to CoA/Spec Sheet.

General, literature-based guidance

• Temperature: 12–37 °C. Lower temperatures (12–25 °C) help limit 3′→5′ exonuclease chew-back; 37 °C maximizes rate but can over-digest 3′ overhangs if dNTPs are absent.
• Time: 5–60 min depending on DNA amount and end structure. Start with 10–20 min and titrate.
• Cofactor: 5–10 mM MgCl2. Avoid EDTA; if present from upstream steps, repurify or supplement Mg2+.
• Buffer: 50 mM Tris-HCl pH 7.5–8.0, 50–100 mM NaCl/KCl, 1 mM DTT, optional 0.05–0.2 mg/mL BSA.
• dNTPs: 50–250 µM each for fill-in; omit to favor chew-back. For labeled incorporation, substitute a portion with modified dNTPs as validated.
• Enzyme amount: Follow manufacturer unit definition. A common starting point in literature is 1–5 U per µg DNA for end-polishing, but optimize for fragment size and composition.
• Quenching/stopping: Add EDTA to 10–20 mM final and/or heat at 65–75 °C for 10–20 min. Verify heat inactivation empirically.
• Cleanup: Use silica spin columns or magnetic beads to remove enzyme and salts before ligation or PCR.

Compatibility notes

• Salt sensitivity: High ionic strength can reduce binding/processivity; keep ≤150 mM total monovalent salt during reaction unless empirically justified.
• Additives: PEG or crowding agents can influence ligation readiness post-polishing but are typically omitted during polymerase steps.

Safety & Handling

Item-specific (from Product Data)

• GHS Classification, signal word, pictograms, H-statements: Not specified for this item; refer to SDS.
• Research Use: For research use only.

General laboratory safety (not a substitute for the SDS)

• PPE: Lab coat, disposable nitrile gloves, and safety glasses. Avoid skin/eye contact and inhalation of aerosols.
• Biological nature: Non-pathogenic recombinant/phage-derived enzyme preparation intended for in vitro use; nonetheless handle using standard aseptic and biosafety practices.
• Incompatibilities: Avoid contact with oxidizers, proteases, detergents (SDS), and chelators (EDTA) during use. Do not mix with phenol/chloroform or alcohols in active enzyme solutions.
• First aid (general):
  • Skin: Wash with soap and water.
  • Eyes: Rinse cautiously with water for several minutes; remove contact lenses if present and easy to do; continue rinsing.
  • Inhalation: Move to fresh air.
  • Ingestion: Rinse mouth; seek medical advice.
• Spill response: Absorb small liquid spills with paper towel, clean with suitable disinfectant (e.g., 10% bleach) followed by water.
• Waste: Dispose according to institutional biosafety and local regulations for non-hazardous biological reagents.
• Special risks: Enzyme is proteinaceous and may cause allergic sensitization in susceptible individuals upon repeated exposure; minimize aerosol generation.

Always consult the product-specific SDS for authoritative hazard, toxicological, and exposure-control information.

Solvent Selection

Applicability

• Not a solvent; T4 DNA polymerase is an aqueous enzyme preparation. Solvent selection in the usual organic-chemistry sense does not apply.

Practical guidance for buffer/media compatibility (literature/general)

• Reaction media: Use nuclease-free water and buffered aqueous systems (e.g., Tris-HCl pH 7.5–8.0) with essential divalent cations.
• Essential cofactor: Mg2+ (commonly 5–10 mM MgCl2). Mn2+ can be used to modulate fidelity but is generally avoided for high-accuracy applications.
• Additives that improve performance: 1 mM DTT (maintains cysteines reduced); 0.05–0.2 mg/mL BSA (stabilizes enzyme and reduces surface adsorption); 50–100 mM NaCl or KCl for ionic strength as needed.
• Incompatible components (avoid or minimize):
  • Chelators (EDTA/EGTA) that sequester Mg2+
  • Detergents like SDS; residual CTAB or guanidinium salts from nucleic acid preps
  • Phenol, chloroform, ethanol/isopropanol (carryover from extraction/precipitation)
  • High salt (>200–300 mM) can reduce binding/processivity
• Storage buffer considerations: Many commercial polymerases are supplied in 50% glycerol to stay liquid at −20 °C; mixing high glycerol into reactions can affect ionic strength and activity. Keep final glycerol ≤5–10% unless validated.

When to choose alternative enzymes (selection context)

• For blunt-ending with minimal exonucleolytic chew-back, Klenow (exo−) is preferred.
• For strong 3′→5′ proofreading/blunt creation from 3′ overhangs, T4 DNA polymerase is often superior.

Storage & Reconstitution

Item-specific (from Product Data)

• Storage conditions: Store at −20 °C.
• Handling: Avoid repeated freezing and thawing.
• Shipping: Ice chest + ice pads.

What is not specified for this item

• Stock concentration, buffer composition, presence/percentage of glycerol or BSA, and whether the product is supplied lyophilized or in solution are not specified; refer to CoA/Spec Sheet.

General guidance (literature/best practice)

• If supplied in solution (common for polymerases): Keep on ice during use. Aliquot upon first thaw to single-use volumes to minimize freeze–thaw.
• If supplied with glycerol (e.g., ~50% typical in industry): Store at −20 °C (remains liquid); avoid frost-free freezers. Do not store at −80 °C unless manufacturer indicates compatibility.
• Avoid metal chelators and proteases in storage buffers. Maintain reducing conditions (e.g., 1 mM DTT) if provided by the manufacturer.
• For lyophilized formats: Reconstitute with nuclease-free water or the provided buffer to the specified concentration; mix gently, avoid foaming, and allow 5–10 min to fully dissolve on ice. Exact reconstitution instructions are not specified for this item.
• Stability monitoring: Loss of activity may present as incomplete end-polishing or poor ligation efficiency; include positive controls to track performance over time.

Disposal

• Dispose of unused enzyme and buffers according to institutional guidelines for non-hazardous biological reagents.

Structure & Identity

Item-specific (from Product Data)

• Product: T4 DNA Polymerase (SKU: FP1508928)
• CAS: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES / InChIKey / CID: Not applicable to proteins or not specified for this item; refer to CoA/Spec Sheet.

Literature/general description of structure

• Identity: T4 DNA polymerase is the replicative DNA polymerase from bacteriophage T4 (a large family B polymerase).
• Subunit architecture: Single polypeptide with N-terminal proofreading domain and C-terminal polymerase domain; belongs to the PolB/alpha-like family.
• Functional domains (literature):
  • 3′→5′ exonuclease proofreading domain (Exo I–III motifs)
  • Polymerase palm, fingers, and thumb subdomains for nucleotide addition
• Catalytic mechanism: Two-metal-ion (Mg2+) catalysis typical of DNA polymerases.
• Size (literature): Approx. 110–115 kDa monomeric enzyme; no quaternary structure required for core polymerase activity.
• 2D structural description in words: Elongated globular enzyme in which a palm-shaped C-terminal catalytic region binds primer–template DNA; an N-terminal exonuclease cleft engages 3′ termini for proofreading, connected by a flexible linker allowing DNA shuttling between active sites.

Notes

• Exact amino-acid sequence, tag, source organism/host, and any mutations (e.g., exonuclease-deficient variants) are not specified for this item; refer to CoA/Spec Sheet.

Synthetic Utility

Context

• In synthetic biology and molecular cloning, T4 DNA polymerase is a powerful tool for constructing defined DNA ends and sequences.

Key utilities (literature/general)

• End repair/blunt-ending: Converts heterogeneous fragment ends (5′ or 3′ overhangs) to ligation-ready blunt ends, simplifying vector–insert assembly.
• Controlled chew-back: Precisely trims 3′ overhangs to generate recessed 3′ termini for downstream fill-in with desired sequences, useful in adapter design and library preparation.
• Gap filling: Completes short gaps in DNA constructs or after nuclease treatments, assisting in site-directed mutagenesis and repair assays.
• Label incorporation: Facilitates incorporation of modified dNTPs (e.g., biotin- or fluorescently labeled) during fill-in for immobilization or detection.
• cDNA second-strand synthesis: Extends RNA–DNA hybrids after RNase H nicking, producing ds cDNA for cloning and sequencing library workflows.

Strategic considerations

• Choose T4 DNA polymerase over Klenow when strong proofreading is desired to remove 3′ mismatches and produce high-quality blunt ends.
• To protect desired 3′ ends, maintain dNTPs in excess and shorten incubation time; work at lower temperatures to temper exonuclease.
• For overhang preservation, use exonuclease-deficient polymerases (e.g., Klenow exo−) instead.

Item-specific caveat

• Specific activity (U/µL) and recommended enzyme-to-DNA ratios are not specified for this item; consult the CoA/Spec Sheet for quantitative guidance.

Target Specificity

Applicability

• Not an antibody or affinity reagent; classical “target specificity” (epitope, clone, isotype) does not apply.

Literature/general catalytic specificity

• Substrate: Primer–template duplex DNA; requires a free 3′-OH on the primer strand.
• Sequence preference: No strict sequence specificity; kinetics and fidelity can vary modestly with local sequence context.
• Directionality: 5′→3′ polymerase activity; 3′→5′ exonuclease proofreading.
• Limitation: Cannot initiate synthesis de novo; requires annealed primer. Will degrade 3′ overhangs in the absence of dNTPs.

Item-specific

• No additional targeting or fusion/DNA-binding domains are specified for this item; refer to CoA/Spec Sheet.

Frequently Asked Questions

How should this product be stored?

Store at ?20 °C. Avoid repeated freeze–thaw cycles. Aliquot into single-use volumes before freezing.

What does Suitable for molecular biology mean?

Suitable for molecular biology indicates that the product has been processed and tested for molecular biology and proteomics. Typical specifications include low or undetectable nuclease/protease activity, low endotoxin, controlled microbial limits and high lot-to-lot reproducibility.

What is EnzymoPure??

EnzymoPure? is one of Aladdin Scientific's premium product lines, formulated for high-purity applications across multiple workflows. Compared to standard grades in the same workflow, EnzymoPure? products typically offer tighter specifications, more rigorous QC, and stronger lot-to-lot consistency.

How is this product shipped?

This product ships in an insulated container with ice pads. Unpack on arrival and transfer it to the storage condition stated above.

What documentation is provided?

Available product documentation, including Certificates of Analysis (COA), Safety Data Sheets (SDS), and specification sheets, is shown in the product document area. Document availability and access follow the current site policy.