- Home
- IVD
- By Technology Types
- By Diseases Types
- By Product Types
- Research
- Resource
- Distributors
- Company
| Product Name | Dual-Labeled Hydrolysis Probe (HEX-BHQ1) for Multiplex qPCR |
| Catalog No. | PRIPRO-0011 |
| Description | A dual-labeled fluorescent hydrolysis probe for multiplex qPCR, utilizing 5′-HEX (hexachlorofluorescein) as the reporter fluorophore and 3′-BHQ1 (Black Hole Quencher 1) as the dark quencher. The HEX fluorophore (Ex 535 nm / Em 556 nm) is spectrally well-separated from FAM, making this probe ideal for duplex or multiplex qPCR assays where two targets are detected simultaneously in a single reaction well. BHQ1 efficiently quenches HEX fluorescence through combined FRET and static contact quenching mechanisms when the probe is intact. Taq polymerase 5′→3′ exonuclease activity cleaves the hybridized probe during PCR extension, liberating HEX from BHQ1 and generating a target-proportional fluorescent signal. The probe is HPLC purified to >95% dual-labeled homogeneity. |
| Intended Use | Designed for multiplex qPCR assays requiring simultaneous detection of two targets (duplex with FAM probe), internal control monitoring, reference gene normalization in the same well, and gene expression profiling with dual-color detection. |
| Principle / Technology | FRET/static quenching of HEX by BHQ1; signal generation via Taq polymerase 5′→3′ exonuclease probe cleavage during PCR; HEX channel detection for second target |
| Detection Method | Real-time fluorescence in HEX channel (Ex 535 nm / Em 556 nm) with simultaneous FAM channel monitoring |
| Sample Type | Purified DNA or cDNA; suitable for co-amplification with FAM-labeled probe in same reaction |
| Performance Range / Specifications | Probe 18–30 bases; Tm 65–70 °C; GC 40–60%; spectral cross-talk with FAM channel <5% after color compensation |
| Sensitivity / LOD | Detection of <10 target copies per reaction; linear range comparable to single-color probe (6–7 logs); performance maintained in multiplex format |
| Specificity | Sequence-specific detection; minimal cross-reactivity with co-amplified targets in multiplex panels; BHQ1 provides efficient quenching specific to HEX emission range |
| Reaction Conditions / Protocol | Use at 100–250 nM in duplex qPCR with FAM-BHQ1 probe at matched concentration. Primers at 300–900 nM. Standard cycling conditions with fluorescence acquisition in both FAM and HEX channels. |
| Components / Formulation | Light-protected tube with lyophilized HEX-BHQ1 dual-labeled probe; COA |
| Storage Conditions | –20 °C in dark; protect from ambient light at all stages |
| Shelf Life | 12 months at –20 °C dry; 6 months in solution at –20 °C (dark) |
| Package Specifications | Amber tube with 5–50 nmol yield; custom packaging available |
| Product Form | Lyophilized yellow-orange pellet (HEX dye visible) |
| Quality Control | HPLC ≥95% dual-labeled purity; MS confirmation; quenching efficiency validated; spectral cross-talk assessment provided; endotoxin testing optional. |
| Key Features | Spectrally distinct from FAM for duplex qPCR; BHQ1 dark quencher eliminates background fluorescence; HPLC purified to >95%; MS sequence verification; compatible with all major qPCR platforms with HEX/JOE/VIC channels. |
| Purity | ≥95% dual-labeled by HPLC; unlabeled/single-labeled <5% |
| Concentration | Lyophilized; resuspend to 100 µM stock; yield 15–40 OD260 units at 25 nmol scale |
| Activity / Unit Definition | Quenching efficiency >95% for HEX-BHQ1 pair; fluorescence signal increase >8-fold upon enzymatic cleavage; minimal cross-talk to FAM channel |
| Molecular Weight | Probe sequence MW plus ~590 Da (HEX) and ~547 Da (BHQ1); reported on COA |
| Source / Origin | Synthetic; HEX and BHQ1 phosphoramidite building blocks |
| pH Range / Optimal pH | Stable and functional in qPCR buffer pH 8.0–8.5; compatible pH 6.0–9.0 |
| Shipping Conditions | Ambient temperature in foil-wrapped packaging; stable during transit |
| Expiration Date / Stability | 12 months dry at –20 °C (dark); 6 months in solution at –20 °C (dark) |
| Regulatory / Compliance | For research use only; not for diagnostic applications |
| Compatibility | HEX/JOE/VIC channel on ABI 7500/QuantStudio, Bio-Rad CFX series, Roche LightCycler 480, Qiagen Rotor-Gene Q; verified minimal cross-talk to FAM channel |
| Recommended Buffer System | Standard qPCR buffer; probe compatible with commercial 2X master mixes for multiplex qPCR |
| Application Notes / Precautions | Perform color compensation run before multiplex assay to account for spectral overlap. Balance probe concentrations (typically 100–250 nM each) for equal endpoint fluorescence. Verify no primer-probe interactions in silico before synthesis. |
| Batch-to-Batch Consistency | HPLC profile and quenching efficiency consistent within ±5% and ±2% respectively; MS mass accuracy ±0.05% |
For research use only, not for clinical use.
|
There is no product in your cart. |
Copyright © 2026 Alta DiagnoTech. All rights reserved.
