- Home
- IVD
- By Technology Types
- By Diseases Types
- By Product Types
- Research
- Resource
- Distributors
- Company
| Product Name | Dual-Labeled Hydrolysis Probe (CY5-BHQ2) for Far-Red qPCR Detection |
| Catalog No. | PRIPRO-0013 |
| Description | A dual-labeled hydrolysis probe for qPCR detection in the far-red spectrum, featuring 5′-Cyanine 5 (CY5) reporter and 3′-Black Hole Quencher 2 (BHQ2). CY5 has excitation and emission maxima at approximately 648 nm and 668 nm respectively, in the far-red region where biological autofluorescence is minimal and spectral separation from visible fluorophores (FAM, HEX, ROX) is maximal. This probe enables four- or five-color multiplex qPCR panels. The CY5-BHQ2 pairing benefits from the broad absorption spectrum of BHQ2 (550–650 nm extending to near-IR), which efficiently quenches CY5 through dual FRET and static quenching pathways. HPLC purification ensures >95% dual-labeled product with minimal background. The lyophilized probe is shipped at ambient temperature in light-protective packaging. |
| Intended Use | Designed as the fourth or fifth detection channel in high-order multiplex qPCR assays for applications including respiratory virus panel testing, foodborne pathogen screening, cancer mutation panel analysis, and any scenario requiring simultaneous detection of multiple targets from limited sample material. |
| Principle / Technology | CY5 fluorescence quenched by BHQ2 via FRET and static quenching; TaqMan probe cleavage during PCR separates CY5 from BHQ2, generating far-red fluorescent signal proportional to target amplification |
| Detection Method | Real-time fluorescence in CY5 channel (Ex ~648 nm / Em ~668 nm); multi-channel detection with FAM, HEX, ROX, and CY5 simultaneously |
| Sample Type | Nucleic acid extracts from clinical swabs, food samples, environmental samples, cell lines, and FFPE tissues for multiplex qPCR analysis |
| Performance Range / Specifications | Probe 18–30 bases; Tm 65–70 °C; minimal spectral overlap with shorter-wavelength probes; CY5 emission at 668 nm is baseline-resolved from ROX (605 nm) |
| Sensitivity / LOD | Detects <10 copies per reaction in five-color multiplex format; linear dynamic range maintained at 6 orders of magnitude |
| Specificity | Sequence-specific detection in far-red channel; BHQ2 quencher tailored for long-wavelength fluorophores; negligible quenching of visible-range fluorophores |
| Reaction Conditions / Protocol | Use at 100–250 nM in multiplex qPCR. Standard two-step cycling: 95 °C 15 s denaturation, 60 °C 45 s annealing/extension with CY5 channel acquisition. Full multi-channel detection protocol. |
| Components / Formulation | Light-protected tube with lyophilized CY5-BHQ2 probe; COA supplied |
| Storage Conditions | –20 °C in dark; CY5 is light-sensitive; minimize exposure during handling |
| Shelf Life | 12 months at –20 °C dry (dark); 6 months in solution at –20 °C (dark) |
| Package Specifications | Foil-wrapped tube; 5–50 nmol scale; COA with spectral characterization |
| Product Form | Lyophilized blue to blue-green pellet (CY5 dye color) |
| Quality Control | HPLC ≥95% dual-labeled purity; MALDI-TOF MS; quenching efficiency measurement; fluorescence emission scan (600–750 nm); residual free CY5 <3%. |
| Key Features | Far-red detection minimizes autofluorescence background; ideal fourth/fifth color in multiplex qPCR panels; BHQ2 dark quencher for low background; HPLC purified >95%; MS sequence verification; compatible with instruments having CY5 detection channel. |
| Purity | ≥95% dual-labeled; residual free dye <3%; single-labeled <2% by HPLC |
| Concentration | Lyophilized; resuspend to 100 µM; yield 15–40 OD260 at 25 nmol scale |
| Activity / Unit Definition | Quenching efficiency >95%; fluorescence increase >8-fold upon cleavage; CY5 extinction coefficient ~250,000 M⁻¹cm⁻¹ at 648 nm |
| Molecular Weight | Probe sequence MW plus ~650 Da (CY5) and ~560 Da (BHQ2); accurate mass reported on COA |
| Source / Origin | Synthetic; CY5 and BHQ2 chemical building blocks |
| pH Range / Optimal pH | CY5 fluorescence pH-stable 5.0–9.0; optimal qPCR buffer pH 8.0–8.5 |
| Shipping Conditions | Ambient temperature in light-protective foil packaging |
| Expiration Date / Stability | 12 months dry at –20 °C (dark); 6 months in TE at –20 °C (dark) |
| Regulatory / Compliance | For research use only; not for diagnostic use |
| Compatibility | CY5 channel on Roche LightCycler 480 (channel 640/668), Bio-Rad CFX series (CY5 channel), ABI QuantStudio with CY5 filter set, Qiagen Rotor-Gene Q CY5 channel; compatible with commercial multiplex master mixes |
| Recommended Buffer System | Standard qPCR buffer (Tris-HCl pH 8.0–8.5, KCl, MgCl₂ 3–5 mM); CY5-BHQ2 probe stable in common buffer systems |
| Application Notes / Precautions | CY5 is susceptible to photobleaching; minimize light exposure during handling and avoid prolonged laser excitation. For five-color multiplex, verify channel separation with pure dye standards before assay setup. Aliquot working stocks to avoid freeze-thaw cycles. |
| Batch-to-Batch Consistency | HPLC dual-label purity ≥95%; CY5 emission peak within ±3 nm; quenching efficiency within ±3% across production lots |
For research use only, not for clinical use.
|
There is no product in your cart. |
Copyright © 2026 Alta DiagnoTech. All rights reserved.
