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With the increasing demand and application of oligonucleotide-based diagnostic probes in biomedical research and industrial production, BOC Sciences looks to the global market to provide you with customization and development of dual-labeled probes. Your various dual-labeled probes project needs, such as various modifications or labeling at the 5' end, 3' end, and specified positions are all available at BOC Sciences.
What are Dual Labeled Probes?
Dual-labeled probes, also known as hydrolysis probes, are 20-30 bp DNA or RNA oligonucleotides that are labeled with two different dye groups at the 5' and 3' ends of the probe. Usually the reporter group is located at the 5' end of the dual-labeled probe and the quenching group is located at the 3' end of the dual-labeled probe.
Mechanism of Dual Labeled Probes
- When the probe remains intact, the quenching moiety suppresses the fluorescence emission of the reporter moiety by fluorescence resonance energy transfer (FRET).
- During the annealing or extension phase of PCR, the probe and primer hybridize to the DNA template while the DNA polymerase moves along the template DNA as the primer is extended. When the enzyme extends to the probe hybridization position, the DNA polymerase 5'-3' nucleic acid exonuclease activity cleaves the probe, which releases the reporter group, causing an increase in fluorescence. The measured fluorescence signal is proportional to the amount of target DNA.
Figure.1 The sturcture of dual labeled probes.
Applications of Dual Labeled Probe
Dual-labeled probes have the advantages of high sensitivity and high specificity, thus realizing accurate quantification in the fields of pathogen detection, gene polymorphism research, and gene mutation detection. Moreover, the features of dual-labeled probes make it possible to perform multiplexed assays to detect multiple target sequences simultaneously.
- Real Time PCR: Dual-labeled probes are used as TaqMan probes in real-time PCR to detect and quantify the amplification of specific DNA sequences during PCR by changes in fluorescence signal.
- Gene Expression Analysis: Dual-labeled probes play an important role in qRT-PCR techniques for genomic studies by quantifying the expression levels of specific mRNA molecules.
- Fluorescence In Situ Hybridization (FISH): The fluorescent signal of a dual-labeled probe visualizes the location of the target sequence within a cell or tissue and can be used for FISH detection.
- Mutation Analysis: Through genotyping assays, dual-labeled probes can be used to detect and analyze gene mutations during disease development, which can be used to develop new targets for disease treatment.
BOC Sciences' Custom Dual Labeled Probes
BOC Sciences has continuously optimized oligonucleotide synthesis and modification technologies and procedures to provide customized dual-labeled probes for your single or multiplexed real-time PCR applications. We offer a wide selection of fluorescent probes, or you can inform us of the labeling site and submit the dye for modification, leaving us to complete the dual-labeled probe preparation.
Available 3' Modification Quenchers in BOC Sciences
- BHQ-1 Quencher(Quenching Range: 480-580 nm, Quenching Max: 535 nm)
- BHQ-2 Quencher(Quenching Range: 550-650 nm, Quenching Max: 579 nm)
- BHQ-3 Quencher(Quenching Range: 550-650 nm, Quenching Max: 672 nm)
- BBQ-650 Quencher(Quenching Range: 550-750 nm, Quenching Max: 650 nm)
- ECLIPSE Quencher(Quenching Range: 390-625 nm, Quenching Max: 522 nm)
- DABCYL Quencher(Quenching Range: 380-550 nm, Quenching Max: 452 nm)
- TAMRA Quencher(Quenching Range: 470-560 nm, Quenching Max: 544 nm)
Figure 2. The FRET principle for selection of quencher and reporter.
Available 5' Modification Reporters in BOC Sciences
| ATTO-390(ex: 390 nm, em: 476 nm) | ATTO-425(ex: 439nm, em: 485 nm) | LCCyan500(ex: 439nm, em: 485 nm) |
| 6-FAM(ex: 495 nm, em: 520 nm) | Fluo(ex: 495 nm, em: 520 nm) | FITC(ex: 490 nm, em: 525 nm) |
| ATTO-495(ex: 498 nm, em: 526 nm) | TET(ex: 521 nm, em: 536 nm) | ATT0-520(ex: 517nm, em: 538nm) |
| JOE(ex: 522 nm, em: 548 nm) | Yakima Yellow(ex: 530 nm, em: 549 nm) | HEX(ex: 535 nm, em: 556 nm) |
| ATTO-Rho6G(ex: 533 nm, em: 557 nm) | Cy3(ex: 546 nm, em: 563 nm) | TAMRA(ex: 564 nm, em: 579 nm) |
| ROX(ex: 576 nm, em: 601 nm) | Texas Red(ex: 586 nm, em: 610 nm) | LCRed610(ex: 590 nm, em: 610 nm) |
| ATTO-Rho13(ex: 603 nm, em: 627 nm) | DY480XL(ex: 500 nm, em: 630 nm) | ATTO-Rho14(ex: 625 nm, em: 646 nm) |
| LCRed640(ex: 625 nm, em: 640 nm) | Cy5.5(ex: 683 nm, em:705 nm) | IRD700(ex: 685 nm, em: 705 nm) |
Features of BOC Sciences' Custom Dual Labeled Probes Services
- GMP-grade custom synthesis services
- Continuously optimized deprotection and purification process
- Allowing for late-stage modifications
- Greatly improved thermal stability, hybridization specificity and assay sensitivity
- Average internal turnaround time: 3-5 business days
- Wide range of applications, recommended for PCR, SNP detection, allele differentiation, sequencing and hybridization, etc.
- Achieve large scale and high throughput
If you have any other oligonucleotide modification requirements, please feel free to contact us.
* Only for research. Not suitable for any diagnostic or therapeutic use.
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