Taqman probe qPCR Method

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TaqMan Probe qPCR Method

The Taqman method depends on a DNA-based probe with a fluorescent reporter at one end and a quencher of fluorescence at opposite end of the probe. The close proximity of the reporter to the quencher prevents emission of its fluorescence, hydrolyzation of the probe by the 5' to 3' exonuclease activity of the Taq polymerase releases the reporter and thus allows emission of fluorescence. Therefore an increase of the product targeted by the reporter probe at each PCR cycle causes a proportional increase of fluorescence. Fluorescence is detected and measured in a real-time PCR machine.

Basic principle of SYBR Green dye method

Amplification curves of target gene in standards and samples

Taqman probe qPCR Applications

Qualitiative Analysis Relative Quantitative Analysis
Absolute Quantilative Analysis SNP Analysis
Mutiplex real-time qPCR Gene Copy Determination

Probe Types and Fluorophores

There are different probe types and fluorophore tags.You can customize probes with different fluorophores according to their demands. You also can choose one or several fluorophores to perform single or multiplex quantitative PCR.

Probe types and fluorophore tags

Parameters of excitation and emission wavelength of fluorophores

qEASY qPCR Primer Pairs and Probes

A successful qPCR assay requires efficient and specific amplification of the product. Both the primers and the target sequence can affect amplification efficiency and specificity. The use of PCR primers specifically designed and validated for qPCR assays with your target of interest is highly recommended. Sinobiological provide ready-to-use qEASY qPCR primer pairs and probes.

qEASY qPCR Primer Pairs are designed using SBI's proprietary primer design algorithm. They are used for SYBR Green dye-based real-time PCR and designed according to the conserved region of all variants of a specific gene. At least one primer crosses the junction of adjacent exons to avoid amplification of genomic DNA directly and effectively. Our primer pairs cover all genes from human, mouse, rat and can be widely applied to the quantitative analysis of gene expression. cDNA used as templates, a single, correct-size band is produced in SYBR Green dye-based PCR with each pair of primers. Therefore, our primers have the characteristics with high specificity, high amplification efficiency, wide linear range and uniform reaction conditions.

qPCR primer pairs qPCR taqman probes
Unique primer design Strong conservative property
Strict validation process High specificity
Uniform PCR conditions to save time and cost High sensitivity
~100% amplification efficiency to ensure the accuracy of quantification Strict validation process
  Probes of high quality

Taqman Probe qPCR Service Process

Taqman Probe qPCR Service Time

A normal qPCR experiment is completed in 2-3 weeks. Time of particular cases are in line with genes and samples detected.

Taqman probe qPCR Service Charge

Please consult our company for detailed information.

Example

Detect the copy number changes of nodavirus after it infect the HighFive cells by absolute quantification qPCR.

Sample description:

Detect the copy numbers after the nodavirus infect HighFive cells for 4 days and 10 days.

Sample names: noda-HF(control), noda-HF(4d), noda-HF(10d).

Detected genes: noda-RNA2

Detection method: Absolute quantification qPCR (TaqMan probe method)

Test experiment report:
1. Standard curves of Nodavirus RNA2

Amplification curves of RNA2

Standard curve of RNA2

2. Report charts of original data for detected samples

Amplification curves of RNA2 in samples

3. Analysis report of test results

Analysis result of the copy numbers of nodavirus in HighFive cells

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Real-time PCR Service: Gene Expression Analysis by qPCR+
- SYBR Green qPCR Method
- Taqman probe qPCR Method
- Qualitative Analysis Service of Gene Expression by qPCR
- Relative Quantitative Analysis Service of Gene Expression by qPCR
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