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Scorpions® Primers and Probes

Self-reporting real-time PCR probes that combine primer and probe function into a single molecule for rapid, highly specific fluorescence detection.

real-time PCR SNP genotyping mutation detection low background signal fluorophore–quencher design HPLC / PAGE purification

What Are Scorpions® Primers?

Scorpions® primers are intramolecular qPCR probes designed for fast signal kinetics, strong allele discrimination, and low background fluorescence

Primers are specialized self-reporting real-time PCR probes that integrate a PCR primer and probe sequence into a single oligonucleotide. The probe region is typically held in a stem-loop structure containing a fluorophore and quencher, while a PCR-blocking moiety prevents polymerase extension into the probe region.

After primer extension, the probe sequence hybridizes back to the newly synthesized target strand within the same molecule. This intramolecular probing mechanism separates the fluorophore from the quencher and produces fluorescence signal during amplification.

Because the probe is physically linked to the primer, Scorpions® technology can provide rapid signal generation, high specificity, and low background fluorescence for demanding qPCR and genotyping workflows.

Scorpions® Mechanism: Intramolecular Real-Time PCR Detection

Scorpions primer intramolecular mechanism showing hairpin, primer extension, probe hybridization, and fluorescence signal generation

Scorpions® intramolecular mechanism. The probe is linked to the primer, enabling rapid hybridization to the newly synthesized strand and fast fluorescence generation without probe diffusion or cleavage.

Scorpions® Primer Services & Pricing

Bio-Synthesis offers fixed-price Scorpions® primer/probe synthesis for real-time PCR and genotyping applications. Scorpions® primers include a DNA probing sequence held in a stem-loop conformation with a 5′ reporter dye, an internal quencher, and an HEG blocker linking the probe to the PCR primer.

Standard Scorpions® primers are HPLC purified, QC checked by MALDI-TOF MS and analytical HPLC, delivered lyophilized, and are typically supplied in 4–6 working days.

Guaranteed Delivered Quantity

Guaranteed OD260 Estimated nmol Estimated µg Estimated Reactions Maximum Length
1 OD 2 nmol 32 µg 400 reactions 60 mer
5 OD 10 nmol 160 µg 2,000 reactions 60 mer
10 OD 20 nmol 320 µg 4,000 reactions 60 mer
20 OD 40 nmol 640 µg 8,000 reactions 60 mer

*Includes dual HPLC purification and mass check.

Uni-Probe Dye, Quencher, and Blocker Options

5′ Fluorophore Options Internal Quencher Blocker
6-FAM™, TET, JOE, HEX, Cy3, TAMRA, Cy3.5, ROX, Texas Red Dabcyl, dT HEG
6-FAM™, HEX™, TET™, JOE™, Oregon Green 514, Bodipy R6G-X, Cy3, Rhodamine Red-X, TAMRA BHQ1 HEG
HEX, TAMRA™, Texas Red®, ROX™, Cy3, Cy3.5, Texas Red-X, Bodipy TR-X, LightCycler 630, Rhodamine Red™, Cy5 BHQ1 HEG
Cy5, Cy5.5 BHQ3 HEG

Some reporter dyes may quench with less-than-optimal efficiency but can still be used in properly designed hairpin probes.

Need probe design, larger molar amounts, large-volume orders, or special dye/quencher combinations?
Bio-Synthesis can help configure Scorpions® primers for your assay format and target sequence.

Scorpions® vs. TaqMan® vs. Molecular Beacons

Scorpions® primers are commonly compared with TaqMan® probes and Molecular Beacons in real-time PCR applications. The key difference lies in the detection mechanism, which directly impacts signal speed, specificity, and assay performance.

Feature Scorpions® TaqMan® Probes Molecular Beacons
Mechanism Intramolecular Cleavage-based Hybridization
Signal Speed ★★★★★ Fastest ★★★☆☆ Moderate ★★★☆☆ Moderate
SNP Specificity ★★★★★ Excellent ★★★☆☆ Good ★★★★★ Excellent
Background Signal ★★★★★ Very Low ★★★☆☆ Moderate ★★★★☆ Low
Assay Complexity ★★★☆☆ Medium ★★☆☆☆ Low ★★★☆☆ Medium
Best Use Cases SNP genotyping, mutation detection, allele discrimination Gene expression, pathogen detection, quantification SNP genotyping, allele discrimination, miRNA detection
Key takeaway: Scorpions® primers provide the fastest signal generation and strongest performance for SNP and mutation detection, while maintaining very low background signal.

Why Use Scorpions® Primers?

Faster Signal Generation

Intramolecular hybridization reduces probe diffusion limitations and produces fluorescence earlier in PCR cycles.

High Specificity

Excellent for SNP genotyping, mutation detection, and allele discrimination assays.

Low Background Signal

The hairpin structure keeps fluorophore and quencher close until target-dependent activation.

Single-Molecule Design

Primer and probe are integrated into one molecule, simplifying assay design.

Robust qPCR Performance

Reliable for low-copy targets, rapid cycling, and high-sensitivity applications.

Customizable Chemistry

Flexible fluorophores, quenchers, blockers, purification, and QC options tailored to your assay.

Scorpions® Primer Design Considerations

Successful Scorpions® probe performance depends on careful coordination between primer design, probe sequence, stem-loop stability, fluorophore–quencher pairing, and PCR-blocking chemistry.

Key Design Factors

  • Optimize stem-loop stability for low background and efficient signal generation
  • Balance primer melting temperature and probe-target melting temperature
  • Place the probe sequence for maximum mismatch discrimination when needed
  • Select fluorophore and quencher combinations matched to the instrument channel
  • Use PCR-blocking chemistry to prevent polymerase read-through into the probe

Information Needed for Design

  • Target sequence and amplicon region
  • Desired mutation, SNP, or target site
  • qPCR instrument and detection channels
  • Preferred fluorophore and quencher, if known
  • Assay type: genotyping, detection, quantification, or screening
Design support: Bio-Synthesis can help evaluate Scorpions® primer design requirements, fluorophore–quencher options, purification needs, and QC strategy.

Custom Synthesis, Purification, and QC

Bio-Synthesis provides custom Scorpions® primer and probe synthesis with options for fluorophore–quencher selection, PCR blocker incorporation, purification, and analytical QC documentation.

Custom Probe Construction

Single-molecule Scorpions® designs incorporating primer, probe, stem-loop, fluorophore, quencher, and blocker elements.

Labeling Options

Common fluorophore and quencher combinations can be selected based on assay platform and spectral requirements.

Purification Options

HPLC, PAGE, or custom purification strategies can be selected based on length, label complexity, and application.

Analytical QC

Mass confirmation, HPLC purity, and supporting analytical documentation can be provided depending on project scope.

Research to Diagnostic Support

Support for research, assay development, validation lots, and recurring production requirements.

Technical Consultation

Discuss design, feasibility, fluorescence channels, synthesis complexity, and delivery requirements with our team.

FAQ

What are Scorpions® primers?

Scorpions® primers are self-reporting real-time PCR probes that combine a PCR primer and probe sequence into one molecule. They generate fluorescence after primer extension when the probe hybridizes intramolecularly to the new target strand.

How are Scorpions® different from TaqMan® probes?

Scorpions® primers use intramolecular hybridization, while TaqMan® probes are separate probes that generate signal through nuclease cleavage during PCR.

When should I use Scorpions® instead of TaqMan®?

Scorpions® are useful when rapid signal generation, low background, and strong mismatch discrimination are important, such as SNP genotyping, mutation detection, and allele discrimination.

Are Scorpions® primers good for SNP genotyping?

Yes. Scorpions® designs are well suited for SNP genotyping because the probe can be positioned to strongly discriminate matched and mismatched targets.

Can Bio-Synthesis make custom Scorpions® primers?

Yes. Bio-Synthesis provides custom Scorpions® primer synthesis with fluorophore, quencher, PCR blocker, purification, and QC options.

What information is needed for a quote?

Send the target sequence, desired primer/probe design if available, mutation or SNP position, fluorophore/quencher preference, purification requirement, quantity, and intended application.

Contact & Quote Request

For the fastest review, send your target sequence, desired Scorpions® design, mutation or SNP position, fluorophore/quencher requirements, quantity, purification preference, and intended application.

Fast quote checklist

  • Target sequence and amplicon region
  • SNP, mutation, or target-discrimination requirement
  • Fluorophore and quencher preference
  • Purification level and quantity
  • Research, screening, diagnostic-development, or recurring-use requirement

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