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| Product Name | Bst DNA Polymerase Large Fragment for Isothermal Amplification |
| Catalog No. | IEC-HMM-0065 |
| Description | The large fragment of Bacillus stearothermophilus DNA polymerase I, possessing 5′→3′ polymerase activity and strong strand displacement capability but lacking 5′→3′ exonuclease activity. This enzyme is the core component for loop-mediated isothermal amplification and other isothermal nucleic acid amplification technologies. Its optimal operating temperature of 60–65 °C enables rapid, specific amplification in a single-temperature incubation without thermal cycling equipment. |
| Intended Use | Isothermal DNA amplification via loop-mediated and strand displacement amplification methods for rapid nucleic acid detection. |
| Principle / Technology | Strand displacement DNA synthesis at constant elevated temperature |
| Detection Method | Real-time fluorescence, turbidity monitoring, or lateral flow detection of amplification products |
| Sample Type | Purified DNA from clinical, environmental, and research specimens |
| Performance Range / Specifications | Processivity: extends approximately 1000 nucleotides per binding event; optimum temperature: 65 °C |
| Sensitivity / LOD | Amplification detectable within 15–30 min at constant 65 °C from as few as 10 copies of target |
| Specificity | Strand displacement activity enables primer-annealing without thermal denaturation; lacks 5′→3′ exonuclease for clean LAMP product formation |
| Reaction Conditions / Protocol | Standard LAMP reaction: 1× isothermal buffer, 0.32 U/µL Bst polymerase, 0.2–1.6 µM each primer, 0.2–1.4 mM each dNTP, 4–8 mM MgSO₄, 65 °C for 30–60 min |
| Components / Formulation | Bst DNA Polymerase Large Fragment (8 U/µL in glycerol storage buffer), 10× isothermal amplification buffer, 100 mM MgSO₄ solution |
| Storage Conditions | –20 °C |
| Shelf Life | 24 months at –20 °C |
| Package Specifications | 8000 U, 32000 U |
| Product Form | Liquid enzyme in glycerol storage buffer |
| Quality Control | Activity: 1 unit incorporates 10 nmol dNTP into acid-insoluble material in 30 min at 65 °C; nuclease contamination: none detected after 16 hr incubation of supercoiled plasmid DNA |
| Key Features | Robust strand displacement activity in typical LAMP primer concentrations makes it the standard choice for isothermal diagnostic assay development |
| Purity | ≥95% by SDS-PAGE for enzymes; ≥98% by HPLC for coenzymes |
| Concentration | Enzymes at specified U/µL; coenzymes at specified mM concentration |
| Activity / Unit Definition | One unit incorporates 10 nmol dNTP into acid-insoluble form in 30 min at specified temperature (polymerases); other enzyme units per product specification |
| Molecular Weight | As specified per enzyme: Taq ~94 kDa; M-MLV RT ~75 kDa; T4 ligase ~55 kDa; coenzymes as labeled |
| Source / Origin | Recombinant, expressed in E. coli; purified by multi-step chromatography |
| pH Range / Optimal pH | pH 7.5–8.5 for polymerase activity; pH 7.5–8.0 for ligation; pH 7.5–8.0 for transcription |
| Shipping Conditions | Dry ice for enzyme shipments; cold packs for coenzyme solutions |
| Expiration Date / Stability | 24 months at –20 °C for enzymes; 12–36 months for coenzymes per product type |
| Regulatory / Compliance | Research use; ISO 9001 manufacturing; GMP-grade options available for diagnostic manufacturing |
| Compatibility | Compatible with standard PCR, RT-PCR, isothermal amplification, and molecular cloning workflows |
| Recommended Buffer System | Tris-HCl, KCl, MgCl₂-based buffers; glycerol-containing storage buffers |
| Application Notes / Precautions | Aliquot enzymes to avoid repeated freeze-thaw; use dedicated nuclease-free workspace; verify enzyme activity for critical applications |
| Batch-to-Batch Consistency | Enzyme specific activity within ±15% of reference lot; functional testing per lot |
For research use only, not for clinical use.
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