Dual-Labeled Probes

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

Read more about

• Applications of Dual-Labeled Probes
• Benefits Of Using Dual-Labeled Probes
• Add Locked Nucleic Acid or MGB:EDQ to Your Probe
  
• How Dual-Labeled Probes Work
• Features of Dual-labeled Probes
• Spectral Properties Table
• Shipping Schedule
• Related Products

Applications of Dual-Labeled Probes

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

Perfect for validating MISSION^® siRNA and shRNA knockdowns.

BENEFITS OF USING DUAL-LABELED PROBES

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

ADD LOCKED NUCLEIC ACID OR MGB:EDQ TO YOUR PROBE

Benefits of Locked Nucleic Acid with BHQ™

• Increased thermal stability and hybridization specificity
• Greater accuracy in SNP detection, allele discrimination, and in vitro quantification or detection
• Easier and more sensitive probe designs for problematic target sequences 
• Has no native fluorescence (emits heat instead of light), resulting in lower background fluorescence
• Increased signal-to-noise ratio, providing higher sensitivity
• Enables a wider choice of reporters for multiplexing

Benefits of MGB:EDQ

• Increased specificity of the probe-target hybridization, reducing the likelihood of non-specific binding
• Higher sensitivity enables the use of shorter probes, which increases the sensitivity of qPCR assays
• Melting temperature adjustment allows for Tm adjustment of the probe, which can be beneficial for designing qPCR probes with optimal Tm values
• Reduced background signal contributes to a reduction in background fluorescence, leading to an improved S:N or signal-to-noise ratio
• Enhanced stability can improve the stability of the probe-target hybrid, reducing the likelihood of probe degradation
• Design flexibility expands the options for probe design, facilitating the creation of efficient qPCR assays
• Enhanced SNP detection via mismatch discrimination

How Dual-Labeled Probes Work

A Dual-Labeled Probe is a single-stranded oligonucleotide labeled with two different dyes. A reporter dye is located at the 5’ end and a quencher molecule located at the 3’ end. The quencher molecule inhibits the natural fluorescence emission of the reporter by fluorescence resonance energy transfer (FRET). The illustration below depicts the mechanism. 

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.

Features of Dual-Labeled Probes

• Amounts: 1, 3, 5, and 10 OD
• Purification: HPLC
• Sequence Lengths: 15 - 40 bases
• Quality Control: 100% mass spectrometry
• Format: Supplied dry in amber tubes
• Custom formats available (normalization, special plates, etc.)

Our probes are provided in a format to simplify your experimental planning.

Guaranteed Yields

*Estimate is based on 4 nmoles or 32 µg for 1 OD and 200 nM in 25 µL reaction (5.0 pmol/reaction). Estimate is based on an average sequence length of 25 bases.

Guaranteed OD Yield	Approx. No. of nmoles	Approx. No. of µg	Approx. No. of Reactions*
1	4	32	800
3	12	96	2,400
5	20	160	4,000
10	40	320	8,000

Spectral Properties Table

Dye	Max. EX (nm)	Max. EM (nm)	Compatible Quencher
6-FAM™	495	520	BHQ™-1, TAMRA™, MGB:EDQ
JOE™	529	555	BHQ™-1, TAMRA™
TET™	521	536	BHQ™-1, TAMRA™, MGB:EDQ
Cal Fluor® Gold 5401	522	544	BHQ™-1
HEX™2	535	556	BHQ™-1, TAMRA™, MGB:EDQ
Cal Fluor® Orange 5602	538	559	BHQ™-1
TAMRA™	557	583	BHQ™-2
Cyanine 3	549	566	BHQ™-2
Quasar® 5703	548	566	BHQ™-2
Cal Fluor® Red 5904	569	591	BHQ™-2
ROX™	586	610	BHQ™-2
TxRd (Sulforhodamine 101-X)	597	616	BHQ™-2
Cyanine 5	646	669	BHQ™-3
Quasar® 6705	647	670	BHQ™-3
Cyanine 5.5	675	694	BHQ™-3

¹JOE/TET alternative
²VIC^® alternative
³Cyanine 3 alternative
⁴TAMRA™ alternative
⁵Cyanine 5 alternative

Shipping Schedule

*Delivery times may be longer due to international transit, customs clearance delays, etc. Large projects will be placed on a delivery schedule based upon project needs.

Quantity (OD)	1	3	5	10
Dual-Labeled Probes with Standard Dyes	3-5 days	3-5 days	3-5 days	3-5 days
Dual-Labeled Locked Nucleic Acid Probes with Standard Dyes	7 days	7 days	7 days	7 days
Dual-Labeled MGB:EDQ Probes with 6’-FAM™, HEX™, or TET™ (inquire for other dyes)	N/A	10-12 days	10-12 days	10-12 days
Dual-Labeled Probes with Post-Synthesis Dyes, with/without Locked Nucleic Acid	7-10 days	7-10 days	7-10 days	7-10 days