Custom TaqMan Probe Synthesis

BOC Sciences offers custom TaqMan probes labeled with various fluorophores and quenchers.

What is TaqMan Probe

• Principle of TaqMan experiment

The TaqMan probe is an oligonucleotide probe designed to pair the target sequence's upstream and downstream primer. A specific fluorescent probe is added along with a pair of primers during PCR amplification, labeled with a reporter fluorophore at the 5' end and a quenched fluorophore at the 3' end. Mechanistically, the fluorescent signal emitted by the reporter group is absorbed by the quenched group when the probe is intact. During PCR amplification, the probe is annealed explicitly to the inner region of the PCR product between the forward and reverse primers. Then 5'-3' exonuclease activity of the Taq enzyme degrades the probe, separating the fluorescent reporter group from the quenched fluorescent group so that the fluorescence monitoring system can receive the fluorescent signal. In other words, every time a DNA strand is amplified, a fluorescent molecule is formed to achieve synchronization between fluorescent signals accumulation and PCR products formation.

Fig.1 Diagrammatic illustration of the TaqMan chemistry. (Gangisetty O, 2009)

Common types of fluorescence:

1. Reported fluorescence: FAM, HEX, JOE, TET, VIC, NED, CY3, Texas Red, CT5
2. Quenched fluorescence: TAMARA, BHQ, MGB-NFQ
3. Reference fluorescence: ROX

• TaqMan Probe Applications

1. Real-time quantitative PCR
2. DNA copy number measurement
3. Bacterial identification assays
4. SNP genotyping
5. Validation of microarray results
6. Pharmacogenomics
7. HLA Genotyping
8. Determination of viral load in clinical specimens (HIV, hepatitis)
9. Bacterial identification assays

TaqMan Probe Synthesis Service

• TaqMan Probe Synthesis Service Content

1. Design: Design the sequence and length of the TaqMan probe based on the target sequence or reference sequence provided by the customer.
2. Synthesis: TaqMan probes are synthesized by solid-phase synthesis using synthetic chemistry.
3. Purification: TaqMan probes are purified by HPLC or PAGE, etc. to ensure their purity and quality.
4. Sequencing: Sequencing of TaqMan probes to ensure their correct sequence.
5. Quantification: TaqMan probes are quantified using methods such as colorimetric or fluorometric methods to ensure their concentration and purity.
6. Packaging and shipping: TaqMan probes are packed in appropriate containers and shipped in a professional manner to ensure their stability and integrity.

In addition, BOC Sciences can provide personalized TaqMan probe synthesis services according to customer needs, and analyze and evaluate the properties and structure of TaqMan probes. Please contact us for more information.

• TaqMan Probe Synthesis Service Features

1. Customers only need to provide the GeneBank registration number for the gene of interest.
2. Free primer and probe design services.
3. Free reaction system optimization services.
4. Free technical support services for experimental process.

• Features of TaqMan Probes

1. High template specificity: Fluorescent probe only binds to the target sequence. Theoretically, the probe method's fluorescent signal only comes from the target sequence and is not affected by non-specific products. Therefore, the specificity of the TaqMan probe qPCR method acquires high specificity and accurate quantification.
2. Multiplex PCR detection: Different fluorescent groups can be labeled for multiple gene amplification and detection in one tube.
3. High sensitivity: During low target genes concentration, the dye method can detect up to 5000 copies for gene quantification, and the exclusively specific amplification signal affords reliable results.
4. RT enzyme, Taq enzyme, miRNA standard, reverse transcription primer, characteristic quantitative primer, probe solution, other reagents, and quality inspection report.

Frequently Asked Questions (FAQ)

What is a TaqMan probe and how does it function in qPCR?

A TaqMan probe is an oligonucleotide labeled with a 5' reporter fluorophore and a 3' quencher. It emits fluorescence only after hybridizing to the target sequence and cleavage by Taq polymerase during PCR, enabling real-time detection.

How are TaqMan probes customized for specific genes?

BOC Sciences designs probes based on target sequences or GeneBank IDs, adjusting probe length, fluorophore/quencher choice, and sequence composition to maximize specificity and sensitivity.

What fluorophore and quencher combinations are available?

Common reporter fluorophores include FAM, HEX, JOE, TET, VIC, NED, Cy3, and Texas Red. Quenchers include TAMRA, BHQ, and MGB-NFQ, with optional reference dyes like ROX for multiplex assays.

How does probe design impact detection sensitivity?

Optimized probe sequences and proper fluorophore/quencher placement ensure high template specificity, minimal background, and accurate quantification even at low target concentrations.

Can TaqMan probes be used for multiplex PCR?

Yes, probes labeled with different fluorophores enable simultaneous detection of multiple target sequences in a single reaction, improving efficiency and reducing assay time.