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| Product Name | SP6 RNA Polymerase |
| Catalog No. | IEC-HMM-0068 |
| Description | A DNA-dependent RNA polymerase purified from E. coli expressing the cloned bacteriophage SP6 RNA polymerase gene. The enzyme exhibits stringent promoter specificity for the SP6 promoter sequence, initiating transcription exclusively downstream of this 18-base pair consensus element. SP6 polymerase synthesizes RNA transcripts at high yield from linearized plasmid templates, producing homogeneous RNA suitable for in vitro translation, RNA probe generation, and structural studies. |
| Intended Use | In vitro synthesis of RNA transcripts from DNA templates containing the SP6 promoter for probe generation, structural RNA production, and expression studies. |
| Principle / Technology | Promoter-specific DNA-dependent RNA synthesis |
| Detection Method | Gel electrophoresis, UV spectrophotometry, or incorporation of labeled NTPs |
| Sample Type | Linearized plasmid DNA or PCR product containing the SP6 promoter sequence upstream of target sequence |
| Performance Range / Specifications | Typical yield: 50–100 µg RNA per µg template in 2 hr at 37 °C; transcript length up to 10 kb |
| Sensitivity / LOD | RNA product detectable within 15 min of reaction initiation |
| Specificity | Strict SP6 promoter specificity; no transcription from T7 or T3 promoters |
| Reaction Conditions / Protocol | 1× SP6 reaction buffer, 0.5 mM each NTP, 0.5–1 µg linearized template DNA, 1 U/µL SP6 polymerase, 37 °C for 1–2 hr; optional DNase I treatment to remove template |
| Components / Formulation | SP6 RNA Polymerase (20 U/µL), 5× SP6 transcription buffer (200 mM Tris-HCl, 30 mM MgCl₂, 10 mM spermidine, 50 mM DTT, pH 7.9), NTP mix (25 mM each), nuclease-free water |
| Storage Conditions | –20 °C |
| Shelf Life | 24 months at –20 °C |
| Package Specifications | 5000 U, 25000 U |
| Product Form | Liquid enzyme in glycerol storage buffer |
| Quality Control | Transcriptional activity verified by incorporation of [³H]-UTP into acid-insoluble RNA; DNase and RNase contamination negative |
| Key Features | Stringent promoter specificity eliminates off-target transcription from related phage promoters in multi-promoter plasmid constructs |
| 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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