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| Product Name | Phi29 DNA Polymerase for Whole Genome Amplification |
| Catalog No. | NATR-HMM-0145 |
| Description | A highly processive DNA polymerase purified from Bacillus subtilis phage phi29 for whole genome amplification from minute quantities of DNA. The enzyme couples robust strand displacement activity with exceptional processivity, synthesizing DNA fragments exceeding 70 kb without dissociating from the template. Combined with random hexamer primers, phi29 polymerase enables isothermal multiple displacement amplification yielding micrograms of high-fidelity DNA from picogram input quantities. |
| Intended Use | Isothermal whole genome amplification from limited or degraded DNA samples for downstream genotyping, sequencing, and archiving. |
| Principle / Technology | Multiple displacement amplification using phi29 DNA polymerase and random hexamer primers |
| Detection Method | Fluorometric DNA quantification; agarose gel electrophoresis; downstream qPCR or sequencing |
| Sample Type | Genomic DNA from single cells, FFPE tissue, dried blood spots, forensic samples, and purified microbial DNA |
| Performance Range / Specifications | Typical DNA yield: 10–40 µg from 10 ng input genomic DNA in 8 hr; amplification bias ≤3-fold across genome |
| Sensitivity / LOD | Reliable amplification from as little as 10 pg of genomic DNA |
| Specificity | Phi29 polymerase exhibits 3′→5′ exonuclease proofreading activity; error rate approximately 1 × 10⁻⁶ |
| Reaction Conditions / Protocol | Sample denaturation: 3 min at 95 °C with random hexamer primers; amplification: 8–16 hr at 30 °C; enzyme inactivation: 10 min at 65 °C |
| Components / Formulation | Phi29 DNA polymerase (10 U/µL), 10× phi29 reaction buffer, random hexamer primer solution (1 mM), dNTP mix (25 mM each), nuclease-free water |
| Storage Conditions | –20 °C for enzyme and dNTPs; –20 °C or room temperature for buffer and primers |
| Shelf Life | 24 months at –20 °C |
| Package Specifications | 100 reactions (10 µL enzyme at 10 U/µL in 50 µL reactions) |
| Product Form | Enzyme: liquid in glycerol storage buffer; other components: liquid solutions |
| Quality Control | Enzyme activity verified by processivity assay; genomic DNA amplification tested with 10 pg, 1 ng, and 10 ng human genomic DNA inputs; amplification success confirmed by qPCR of 8 loci across chromosomes |
| Key Features | Strand displacement activity eliminates the need for thermal cycling; proofreading function preserves sequence integrity |
| Purity | DNase/RNase free; PCR inhibitor free; A260/A280 ≥1.8 for nucleic acid products |
| Concentration | As specified on product label; enzyme units per µL as stated |
| Activity / Unit Definition | Polymerase activity defined as nmol dNTP incorporated per 30 min per unit; other activities per product specification |
| Molecular Weight | As specified per DNA markers or protein components |
| Source / Origin | Recombinant enzymes expressed in E. coli; synthetic oligonucleotides; ultrapure reagents |
| pH Range / Optimal pH | pH 7.5–8.5 for PCR; pH 8.0 for most molecular biology applications |
| Shipping Conditions | Cold packs or dry ice for enzymes; ambient for buffers and DNA markers |
| Expiration Date / Stability | 12–24 months at –20 °C for enzymes; 12–24 months at room temperature for buffers |
| Regulatory / Compliance | Research use; ISO 13485 manufacturing for select products |
| Compatibility | Compatible with standard thermal cyclers, real-time PCR instruments, and gel electrophoresis systems |
| Recommended Buffer System | Tris-HCl, KCl, MgCl₂-based reaction buffer; TE for nucleic acid storage |
| Application Notes / Precautions | Use dedicated PCR workspace; aliquot enzymes to avoid contamination; monitor no-template controls |
| Batch-to-Batch Consistency | Enzyme activity within ±15% of reference lot; PCR performance verified with control template |
For research use only, not for clinical use.
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