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| Product Name | CRISPR-Cas12a Fluorescent Detection Reagent System |
| Catalog No. | NATR-HMM-0146 |
| Description | A nucleic acid detection system utilizing the collateral cleavage activity of Cas12a (Cpf1) enzyme for rapid and specific detection of amplified DNA targets. Upon crRNA-guided target recognition, activated Cas12a non-specifically cleaves nearby single-stranded DNA reporter probes, generating a fluorescent signal proportional to target concentration. This system is compatible with isothermal amplification pre-amplification and provides an orthogonal detection mechanism for diagnostic assay development. |
| Intended Use | Sequence-specific detection of amplified DNA using CRISPR-Cas12a trans-cleavage activity for diagnostic assay development. |
| Principle / Technology | CRISPR-Cas12a collateral cleavage of ssDNA fluorescent reporter |
| Detection Method | Real-time fluorescence (FAM channel, 485/535 nm excitation/emission) |
| Sample Type | Amplified DNA from RPA, LAMP, or PCR reactions; can be adapted for direct detection after isothermal pre-amplification |
| Performance Range / Specifications | Time to result: 5–15 min after adding detection mix to pre-amplified sample |
| Sensitivity / LOD | Detection limit: approximately 0.1 nM target DNA after pre-amplification |
| Specificity | crRNA-programmable specificity; single nucleotide discrimination achievable with optimized crRNA design |
| Reaction Conditions / Protocol | Pre-amplified product: 2 µL added to 18 µL detection mix (Cas12a, crRNA, ssDNA-FQ reporter in reaction buffer); incubate 15 min at 37 °C with fluorescence reads every 30 sec |
| Components / Formulation | Cas12a nuclease (1 µM), 10× Cas12a reaction buffer, ssDNA FQ reporter substrate (FAM-BHQ1, 10 µM), nuclease-free water; crRNA must be designed and ordered separately per target |
| Storage Conditions | –20 °C for enzyme and reporter; protect reporter from light at all times |
| Shelf Life | 12 months at –20 °C |
| Package Specifications | 500 reactions (2 µM Cas12a final in 20 µL reactions) |
| Product Form | Liquid enzyme and reporter in storage buffers |
| Quality Control | Each lot functionally validated using a provided synthetic target DNA and crRNA pair; signal-to-noise ratio ≥20 at 10 min |
| Key Features | Programmable crRNA sequence enables rapid reconfiguration of detection specificity without reformulating core reagents |
| 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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