Dual Labeled Probes

Eurofins offer Hydrolysis probes for TaqMan assays with a variety of dye - quencher combinations compatible with all type of instruments.

Dual-Labeled Probes are the most common probe type for qPCR and are often referred to as hydrolysis probes.

Choose Dual-Labeled Probes for:

• Microarray validation
• Multiply target genes / few samples
• Pathogen detection
• Multiplexing
• Viral load quantification
• Gene expression analysis
• Gene copy determination

Benefits of Using Dual-Labeled Probes include:

• Design simplicity for sequence specificity
• Extensive availability of reporter/quencher combinations
• Increased sensitivity

The primer is elongated by the polymerase and the probe binds to the specific DNA template. Hydrolysis releases the reporter from the probe/target hybrid, causing an increase in fluorescence. The measured fluorescence signal is directly proportional to the amount of target DNA.

Specification of Dual Labelled probes

HPLC purified probes for stronger illuminating power

Product specifications:

• HPLC purification included
• Wobbles (non-defined ratio) possible
• TAT of 7-10 WD for Indian production; 2-3 week from GER/US Site
• Delivery: lyophilised in 2.0mL tube
• Quality control by OD measurement and MALDI-TOF MS

1 The synthesis scale indicates the initial amount of 3'-bases.

Average yield applies to a 20mer; Calculation: 1OD=5nmol=30ug; may vary for sequences with GC content >70%, >3 purine streches, or strong secondary structures.

Principle of dual labeled probes

Dual labeled probes used in quantitative real-time PCR systems take advantage of the 5' -> 3' exonuclease activity of Taq polymerase.

Dual labeled probes contain a 5' fluorescent reporter and a 3' quencher which anneals between the PCR primers.

During the extension phase of PCR, the 5' -> 3' exonuclease activity of Taq polymerase cleaves the fluorescent reporter from the probe. The amount of free reporter accumulates as the number of PCR cycles increases. The fluorescent signal from the free reporter is measured in real time and allows the quantification of the target sequence.